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US

20130004580A1

( 1 2 ) Patent

Application

Publication ( 1 0 )

P u b .

N o . : US 2 0 1 3 / 0 0 0 4 5 8 0 A 1

( 1 9 ) United

S t a t e s

L i n e t a l .

( 4 3 ) P u b . D a t e : Jan.

3 , 2 0 1 3

( 5 4 ) DIVALENT

CATION/TALC CONTAINING P u b l i c a t i o n

C l a s s i ? c a t i o n

COMPOSITIONS

AND METHODS FOR

( 5 1 ) I

t

Cl

11

. .

A61P 7 / 0 0 ( 2 0 0 6 . 0 1 )

_ _

A61P 7/16 (2006.0 1)

( 7 6 ) I n v e n t o r s :

C o n n i e

B .

L i n , B e l l e

M e a d ,

NJ

U S ) ;

A61K35/60

( 2 0 0 6 0 1 )

E‘len Th‘mlas Graham Ekma“ Gun“,

( 5 2 )

us.

l .

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 424/523; 424/683

H o p e w e l l , N J ( U S ) ;

Y a - P i n g

H u ,

( 5 7 )

ABSTRACT

H i g h l a n d P a r k ,

NJ

U S ) ; Neema

K u l k a r n i , R a n d o l p h , N J

( U S ) ; Mary

C a t h e r i n e Mack, L a W r e n c e v i l l e ,

NJ

( U s )

( 2 1 ) A p p 1 . N o . : 1 3 / 1 7 0 , 2 9 7

( 2 2 )

F i l e d : J u n .

2 8 , 2 0 1 1

C o m p o s i t i o n s

a n d

m e t h o d s

f o r t r e a t i n g

a n d / o r p r e v e n t i n g

c o n d i t i o n s

s u c h a s

d i a p e r r a s h and a t o p i c d e r m a t i t i s a r e

d i s

c l o s e d .

The o m p o s i t i o n s

a n d m e t h o d s a r e p a r t i c u l a r l y u s e f u l

i n t h e t r e a t m e n t a n d

p r e v e n t i o n

o f d i a p e r r a s h a n d d i a p e r

d e r m a t i t i s

c a u s e d b y t h e p r o l o n g e d c o n t a c t

o f

human s k i n

W i t h

b o d y W a s t e .

T h e m e t h o d s

e m p l o y t h e

t o p i c a l

a p p l i c a t i o n

of

t r y p s i n - i n h i b i t i n g

a g e n t t o t h e a r e a i n

need

f

such r e a t

m e n t ,

o r t h e a r e a Where p r e v e n t i o n

i s d e s i r e d .

The t r y p s i n

i n h i b i t i n g

a g e n t i s

p r e f e r a b l y

a

s p e c i ? e d d i v a l e n t c a t i o n , i . e . ,

m a g n e s i u m ,

i n

c o m b i n a t i o n

W i t h t a l c .

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P a t e n t A p p l i c a t i o n P u b l i c a t io n J a n . 3 ,

2013

S h e e t 1

0 f 3 US

2 0 1 3 / 0 0 0 4 5 8 0

A 1

F i g u r e

1

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P a t e n t A p p l i c a t i o n P u b l i c a t io n J a n . 3 ,

2013

S h e e t

2 0 f 3 US

2 0 1 3 / 0 0 0 4 5 8 0

A 1

S a l t c o n c e n t r a t i o n

(mM)

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P a t e n t A p p l i c a t i o n P u b l i c a t io n J a n . 3 ,

2013

S h e e t

3

0 f

3 US

2 0 1 3 / 0 0 0 4 5 8 0

A 1

F i g u r e 3

100

P r e p a r a t i n n - ‘ 3 P r e p a r a t i n n - d P r e p a r a t i o n - 5 B a b y P a s t a ? P r e p a r a t i n n - ? P r e p a r a t i n n - Y P r e p a r a t i o n - 8

{ 2 5 9 1 5 3 1 0 } , p H

5

t r y p s n

n h i i t n r yc t i v i t y

)

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US 2013/0004580

A 1

DIVALENT

CATION/TALC

CONTAINING

COMPOSITIONS

AND

METHODS

FOR

TREATING

AND/OR PREVENTING

ENZYMATIC IRRITATION

FIELD

OFTHE

INVENTION

[ 0 0 0 1 ]

The

n v e n t i o n

r e l a t e s t o t h e

u s e o f

d i v a l e n t c a t i o n s t o

p r o t e c t

t h e

s k i n

o f a human. More

a r t i c u l a r l y ,

t h e

i n v e n t i o n

r e l a t e s t o t h e

use of

i v a l e n t c a t i o n s i n a

s k i n c a r e

composition

t h a t

c a n b e t o p i c a l l y

a p p l i e d t o

p r o t e c t s k i n f r o m i r r i t a n t s , a n d

i n

p a r t i c u l a r

p r o t e a s e e n z y m e s , a n d more

i n p a r t i c u l a r

t r y p s i n . The r e s e n t i n v e n t i o n i s u s e f u l i n t h e

p r e v e n t i o n

a n d

t r e a t m e n t

o f

d i a p e r r a s h a n d

t h e

m i t i g a t i o n

o f

a t o p i c d e r m a

t i t i s .

BACKGROUND OF

THE INVENTION

[ 0 0 0 2 ] The s k i n

i s

a n a t u r a l

b a r r i e r t o

t h e p e n e t r a t i o n o f

f o r e i g n s u b s t a n ce s . The

s t r a t u m

corneum

s

t h e s u p e r ? c i a l

c o r n i ? e d

l a y e r

o f

t h e s k i n

t h a t

p r o v i d e s

a

b a r r i e r

t o

W a t e r

e v a p o r a t i o n a n d

r e d u c e s

t h e

p e r m e a t i o n o f u n d e s i r a b l e mol

e c u l e s from t h e e x t e r n a l e n v i r o n m e n t .

The s t r a t u m

corneum

c o n s i s t s

of dead c e l l s

c a l l e d c o m e o c y t e s , Which a r e embed

d e d

i n

a l i p i d - r i c h m a t r i x o f

f a t t y - a c i d s ,

c e r a m i d e s , a n d c h o

l e s t e r o l s .

T h i s s t r u c t u r e

o f

c o m e o c y t e s

embedded

n

l i p i d s i s

t h o u g h t t o p r o v i d e many f

t h e

b a r r i e r p r o p e r t i e s o f t h e s k i n .

S u b s t a n c e s d e p o s i t e d on h e s k i n

must r a v e r s e

t h i s s t r u c t u r e

t o

g a i n a c c e s s

to t h e

u n d e r l y i n g

v i a b l e

l a y e r s o f

t h e

s k i n . S k i n

in?ammation

o c c u r s

W h en u b s t a n c e s Which a r e

i r r i t a t i n g

t o

t h e s k i n a r e

a b l e

t o

p e n e t r a t e

t h i s b a r r i e r and i n i t i a t e t h e

s e c r e t i o n

o f in?ammatory

m e d i a t o r s o n c e t h e y

c o n t a c t t h e

s k i n c e l l s i n t h e v i a b l e e p i d e r m i s

a n d d e r m i s

l a y e r s . As t h e

s k i n b a r r i e r

i s

c o m p r o m i s e d , s k i n

i s

s u b j e c t

t o i n ? a m m a t o r y

e v e n t s f r o m p e r c u t a n e o u s a b s o r p t i o n

o f

i r r i t a n t s t h r o u g h t h e

stratum

corneum.

[ 0 0 0 3 ]

S k i n

b a r r i e r

f u n c t i o n

c a n

b e

compromised

by v a r i

e t y of n s u l t s t h a t

c a u s e

in?ammation. One s u c h i n s u l t c a u s e d

by e x p o s u r e t o body

? u i d s

and a s t e r e s u l t s

i n What

s

com

monly

knoWn a s

d i a p e r r a s h . B o d i l y

? u i d s a n d

W a s t e s

may

c o n t a i n s k i n i r r i t a n t s i n t h e form of enzymes such a s p r o

t e a s e s , u r e a s e s , a n d l i p a s e s . Enzymes f o u n d i n f e c e s c l e a v e

t h e s t r a t u m corneum r o t e i n s

a n d

l i p i d s c a u s i n g t h e

b r e a k

doWn f

t h e n a t u r a l

b a r r i e r o f

t h e

s k i n a n d

r e l e a s e

o f

r r i t a t i n g

a g e n t s

such

a s f r e e f a t t y a c i d s . B a c t e r i a l u r e a s e s on

h e

s k i n

c o n v e r t t h e

u r e a

i n u r i n e t o ammonia e s u l t i n g i n

an

a l k a l i n e

pH on

t h e

s k i n . P r o l o n g e d

e x p o s u r e

o f

t h e

s k i n

to

t h e s e

enzymes s t h o u g h t

t o b e a

major c a u s e

o f s k i n damage t h a t

l e a d s t o d e r m a t i t i s a n d s u b s e q u e n t s k i n b r e a k d o W n .

[ 0 0 0 4 ] A umber o f a p p r o a c h e s a r e knoWn

o r

p r o t e c t i n g

t h e s k i n a g a i n s t t h e a c t i o n

o f

p r o t e a s e s a n d s u b s e q u e n t s k i n

b r e a k d o W n ,

i n c l u d i n g

s k i n

p r o t e c t a n t

f o r m u l a t i o n s ,

a n d

a n t i

i n ? a m m a t o r y c o m p o s i t i o n s . Many

f h e s k i n p r o t e c t a n t f o r

m u l a t i o n s

c o m m e r c i a l l y a v a i l a b l e may o t

p r o v i d e a d eq u a t e

p r o t e c t i o n a g a i n s t s k i n

i r r i t a n t s .

Many f t h e se f o r m u l a t i o n s

c o n s i s t

o f s o f t e n i n g

c r e a m s

a n d

h y d r a t i n g compounds

t o

r e p l e n i s h t h e m o i s t u r e

c o n t e n t

o f t h e s k i n . H o W e v e r , t h e s e

f o r m u l a t i o n s d o n o t

b l o c k

t h e

i r r i t a n t s

p r e s e n t i n u r i n e , f e c e s ,

o r blood

s u c h a s , e . g . ,

t h e

p r o t e o l y t i c enzymes

p r e s e n t

i n

f e c e s . F u r t h e r m o r e t h e s e

c o m p o s i t i o n s

o f t e n

c o n s i s t o f

p e t

r o l a t u m , l a n o l i n o r g r e a s y compounds t h a t

can

r u b

o f f

o n t o

g a r m e n t s

a n d d e c r e a s e t h e

a b s o r b e n c y o f

t h e

g a r m e n t .

[ 0 0 0 5 ] A n o t h e r method

o f

t r e a t i n g s k i n i r r i t a t i o n i n c l u d e s

t h e

u s e o f a n ti - i n ? a m m a t o r y c o m p o u n d s . H o W e v e r ,

t h i s

method

o e s n o t

p r o t e c t

t h e s k i n

from

coming

n

c o n t a c t

W i t h

J a n .

3 ,

2 0 1 3

an i r r i t a n t , t h e r e f o r e , damage t o t h e s k i n s t i l l o c c u r s . The

a n t i - i n ? a m m a t o r y s u b s t a n c e

m i t i g a t e s t h e i n ? a m m a t o r y

r e s p o n s e

b u t i t d o e s

n o t

p r e v e n t t h e s k i n

damage

h a t e l i c i t s

t h e i n ? a m m a t o r y e v e n t

i n t h e

? r s t

p l a c e .

[ 0 0 0 6 ]

Hahn

o f

Cosmederm

T e c h n o l o g i e s

d i s c l o s e s

examples Where i v a l e n t c a t i o n s a t

amounts

o f a b o u t 60

mM

o r

g r e a t e r i n h i b i t

s k i n

i r r i t a t i o n c a u s e d

by

c h e m i c a l

i r r i t a n t s

o r

e n v i r o n m e n t a l

c o n d i t i o n s. S e e ,

e . g . , US. P a t . No.

6 , 4 5 5 ,

0 7 6 ; WO9619182;WO9619181; . S .

P a t . N o .

5 , 9 5 8 , 4 3 6

a n d

US. a t . No. 5 , 8 0 4 , 2 0 3 .

[ 0 0 0 7 ] US. P u b l i s h e d

A p p l i c a t i o n N o .

20040102429 t o

Modak t

a l .

d i s c l o s e s

a n t i - i r r i t a n t c o m p o s i t i o n s t h a t c o n t a i n

tWo o r more a t e r - s o l u b l e o r g a n i c sa lts of i n c a t concentra

t i o n s betWeen 0.1 and 2 .

[ 0 0 0 8 ]

US. P a t . N o . 5 , 9 6 5 , 6 1 0 t o

C o l u m b i a U n i v e r s i t y

d i s c l o s e s z i n c g l u c o n a t e

c o n t a i n i n g

t o p i c a l c o m p o s i t i o n s

Which have an a n t i - i r r i t a n t e f f e c t on s k i n .

[ 0 0 0 9 ]

US. a t . No. 4 , 4 7 7 , 4 3 9 t o D ’ A l e l i o e t a l . d i s c l o s e s

t h e u s e o f

s u l f a t e s

o r

p h o s p h a t e s o f b a r i u m , c a l ci u m , s t r o n

tium

or

z i n c

t o

reduce

i r r i t a t e d

o r excoriated a r e a s

surround

i n g

t h e

s t o m a

o f o s t o m y p a t i e n t s .

[ 0 0 1 0 ] Baby s k i n i s

e s p e c i a l l y

s e n s i t i v e

t o e n v i r o n m e n t a l

a n d d i e t a r y c o n d i t i o n s .

F r e q u e n t e x p o s u r e t o

f e c e s a n d

u r i n e

can promote b a c t e r i a l groWth t h a t i s knoWn t o

c a u s e

s k i n

i r r i t a t i o n and i n f e c t i o n .

Consequent

t o t h i s e x p o s u r e ,

e x c e s

s i v e a m o u n t s o f

l i p a s e

a n d p r o t e o l y t i c e n z y m e s , a n d t h e i r

m e t a b o l i c p r o d u c t s s u c h a s ammonia and

f a t t y

a c i d s , a r e

r e l e a s e d

and c a u s e s k i n i r r i t a t i o n and

a l l e r g i c

s k i n r a s h e s .

[ 0 0 1 1 ]

T h e

s k i n p r o b l e m s

d i s c u s s e d

h e r e i n g e n e r a l l y

e x t e n d

t o

humans t t h e e a r l y a n d a t e p o r t i o n s o f

i f e .

T h a t

s ,

i n f a n t s , a s W e l l a s

t h e

e l d e r l y , a r e

o f t e n

s u b j e c t t o

t h e s e

s k i n

p r o b l e m s .

[ 0 0 1 2 ]

One f t h e

most common o n d i t i o n s

o f

i n f a n t s k i n

i s

d i a p e r

r a s h , a l s o knoWn s

d i a p e r

d e r m a t i t i s .

T h e

p r i m a r y

c o n t r i b u t o r s

t o

t h e

d e v e l o p m e n t o f

d i a p e r r a s h

h a v e

l o n g

b e e n

t h o u g h t

t o b e a g e n t s a n d

e n z y m a t i c

a c t i v i t i e s W i t h i n

i n f a n t

u r i n e

a n d f e c e s .

A t h e r t o n ,

A e v i e W o f

t h e

p a t h o p h y s i o l o g y ,

p r e v e n t i o n

a n d

t r e a t m e n t o f i r r i t a n t d i a p e r

d e r m a t i t i s ,

C u r r .

M e d . R e s .

O p i n . , 2 0 : 6 4 5 - 6 4 9 ( 2 0 0 4 ) ; Wolf

t a l . ,

D i a p e r

d e r m a t i t i s , C l i n . D e r m a t o l . , 1 8 : 6 5 7 - 6 6 0 ( 2 0 0 0 ) .

[ 0 0 1 3 ]

H o W e v e r , m e r e l y k e e p in g t h e a r e a

c l e a n

a n d

d r y

d o e s

n o t p r o t e c t t h e i r r i t a t e d

s k i n

from t h e i r r i t a t i o n a s s o c i

a t e d W i t h t h e

b y - p r o d u c t s o f

i n f a n t u r i n e a n d

f e c e s .

US. a t .

No.

5 , 4 3 6 , 0 0 7 t o Abbott L a b o r a t o r i e s d i s c l o s e s a composi

t i o n

t h a t c o n t a i n s a l i n e a r p o l y d i m e t h y l s i l o x a n e p o l y m e r , a

n o n - i o n i c e m u l s i ? e r

c o n s i s t i n g o f

p o l y o x y e t h y l e n e s o r b i t a n

f a t t y

a c i d e s t e r s ,

s o r b i t a n

f a t t y a c i d

e s t e r s ,

p o l y o x y e t h y l e n e

a l c o h o l s ,

o r p o l y o x y e t h y l e n e f a t t y e t h e r s , a l o e v e r a , a n

a l k o x y l a t e d

e t h e r /

s t e r ,

s o d i u m

c i t r a t e , c i t r i c

a c i d , a

b l e n d

o f

p r o p y l e n e

g l y c o l , d i a z o l i d i n y l u r e a ,

m e t h y l p a r a b e n a n d

p r o

p y l p a r a b e n , f o r u s e i n t h e

p r o t e c t i o n

a n d t r e a t m e n t

o f

d i a p e r

r a s h .

[ 0 0 1 4 ]

G r e a t B r i t a i n

US. a t .

No. 1 , 3 5 7 , 7 3 1

t o Lake

d i s

c l o s e s a

poWder

c o m p o s i t i o n t h a t

c a n

be

i n c o r p o r a t e d

i n t o a

h y d r o p h o b i c

o i n t m e n t . A u f f e r

s y s t e m

i s p r o v i d e d t o b u f f e r

t h e c o m p o s i t i o n a t a pH

f

from 5 . 5 t o 7 . 5 .

[ 0 0 1 5 ]

US. P a t .

No.

4 , 5 5 6 , 5 6 0 to t h e

P r o c t o r &

Gamble

Company

d i s c l o s e s

e x a m p l e s o f t h e

u s e

o f l i p a s e i n h i b i t i n g

a g e n t s

a t

amounts

of

about 10

mM

r g r e a t e r , such

a s

Water

s o l u b l e m e t al l i c

s a l t s

i n c l u d i n g

z i n c

c h l o r i d e ,

i n t h e

t r e a t m e n t

o f d i a p e r r a s h .

[ 0 0 1 6 ]

US. a t .

No. 4 , 8 1 6 , 2 5 4 t o Moss i s c l o s e s a compo

s i t i o n

f o r

t r e a t i n g s k i n

i r r i t a t i o n s

s u c h a s d i a p e r r a s h

t h a t

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US 2013/0004580

A 1

c o n t a i n s

z i n c

o x i d e, b o r i c a c i d , k a r a y a g u m , P e r u v i a n

b a l s a m ,

c o d

l i v e r

o i l ,

s o l v e n t

a n d

a p h a r m a c e u t i c a l c a r r i e r .

[ 0 0 1 7 ] Us. P a t .

No. 4 , 9 1 1 , 9 3 2

t o J o h n s o n

and

J o h n s o n

Consumer P r o d u c t s ,

I n c . d i s c l o s e s a

s k i n

c a r e c o m p o s i t i o n

t h a t

may

e u s e d

f o r

d i a p e r r a s h

t h a t

c o n t a i n s z i n c o x i d e

and

m i c o n a z o l e n i t r a t e .

[ 0 0 1 8 ]

Us.

P u b l i s h e d

A p p l i c a t i o n N o .

20050175719

to

Sun

t

a l . d i s c l o s e s

t h e

use of procyanidin

o

p r e v e n t o r t r e a t

s k i n

r a s h .

[ 0 0 1 9 ] Us. P u b l i s h e d A p p l i c a t i o n

N o .

20070237834

to

Bioderm Research

d i s c l o s e s t h e

use of

i v a l e n t

metal

com

p l e x e s o f e o l i t e s ,

s u c h as

z i n c z e o l i t e , t o t r e a t d i a p e r r a s h .

[ 0 0 2 0 ]

Many

k i n

c a r e p r o d u c t s a i m i n g t o r e d u c e t h e symp

toms o f

o r

to r e l i e v e

d i a p e r

r a s h c o n t a i n z i n c o x i d e , Which

f o r m s

a p r o t e c t i v e b a r r i e r . Common r o d u c t s i n c l u d e d i ap e r

r a s h

a n d

s u n s c r e e n c o m p o u n d s .

A l t h o u g h

t h e f o r m u l a t i o n s

a r e

o f t e n

e f f e c t i v e at

p r o v i d i n g

a b a r r i e r ,

t h e y

a r e

o f t e n

g r e a s y .

Z i n c o x i d e i s i n s o l u b l e a n d a p p e a r s t o b e

i n e r t ,

t h a t i s , i t

appears n o t t o

r e a c t

t o m i t i g a t e an in?ammatory

response

o r

to

r e a c t

W i t h

i r r i t a n t s

t o

n e g a t e

t h e

damage

from

u r i n e o r

f e c e s . Z i n c o x i d e

a l o n e

d o e s

n o t

h a v e s i g n i ? c a n t p r o t e a s e

i n h i b i t i o n

c a p a b i l i t y .

[ 0 0 2 1 ] I t i s

no W

n d e r s t o o d

t h a t

t h e

i n i t i a l

s t a g e s

of

some

t y p e s o f d i a p e r r a s h

a r e

t h e r e s u l t

o f

s k i n

i r r i t a t i o n c a u s e d

by

c o n t a c t W i t h d i g e s t i v e enzymes

p r e s e n t

i n i n f a n t f e c e s , p a r

t i c u l a r l y t r y p s i n ,

c h y m o t r y p s i n

a n d e l a s t a s e .

Among h e s e

e n z y m e s

a r e p r o t e o l y t i c a n d

l i p o l y t i c e n z y m e s

p r o d u c e d i n

t h e

g ast r o i n testi n al

t r a c t t o d i g e s t f o o d . I n i n f a n t s , t h e f e c e s

t e n d

t o

b e W a t e r y a n d c o n t a i n , among

t h e r m a t e r i a l s

s u c h a s

b a c t e r i a , some

amounts

o f

d i g e s t i v e

e n z y m e s .

These

e n z y m e s , i f t h e y

r e m a i n

i n c o n t a c t W i t h t h e s k i n f o r

a n y

a p p r e c i a b l e p e r i o d o f t i m e , h a v e been found

t o

c a u s e

an

i r r i t a t i o n

t h a t

i s u n c o m f o r t a b l e and can r e d i s p o s e t h e s k i n

t o

i n f e c t i o n b y m i c r o o r g a n i s m s .

[ 0 0 2 2 ]

P r o t e a s e s

o c c u r n a t u r a l l y i n

a l l

o r g a n i s m s . T h e s e

enzymes a r e

involved

i n a

multitude of

h y s i o l o g i c a l r e a c

t i o n s . P r o t e a s e s c a n

e i t h e r b r e a k s p e c i ? c p e p t i d e b o n d s

( l i m

i t e d p r o t e o l y s i s ) , d e p e n d i n g on h e amino a c i d

s e q u e n c e o f

a

p r o t e i n ,

o r

b r e a k doWn a c o m p l e t e

p e p t i d e

to amino a c i d s

( u n l i m i t e d p r o t e o l y s i s ) .

[ 0 0 2 3 ] P r o t e a s e s

a r e i n v o l v e d i n

d i g e s t i n g

l o n g

p r o t e i n

c h a i n s

i n t o

s h o r t f r a g m e n t s , s p l i t t i n g t h e p e p t i d e

b o n d s

t h a t

l i n k

amino

a c i d r e s i d u e s .

Some of

them

can

e t a c h

t h e

t e r m i

n a l a m i n o a c i d s f r o m t h e p r o t e i n c h a i n ( e x o p e p t i d a s e s , s u c h

a s a m i n o p e p t i d a s e s ,

c a r b o x y p e p t i d a s e A ) ;

o t h e r s a t t a c k

i n t e r

n a l

p e p t i d e

b o n d s

o f

a p r o t e i n ( e n d o p e p t i d a s e s ,

s u c h

a s

t r y p s i n , c h y m o t r y p s i n ,

p e p s i n , p a p a i n , a n d

e l a s t a s e ) .

[ 0 0 2 4 ] P r o t e a s e s a r e d i v i d e d

i n t o

f o u r major r o u p s

a c c o r d

i n g

to

t h e

c h a r a c t e r

o f

h e i r

c a t a l y t i c

a c t i v e s i t e a n d

c o n d i t i o n s

o f a c t i o n : s e r i n e p r o t e i n a s e s , c y s t e i n e ( t h i o l ) p r o t e i n a s e s ,

a s p a r t i c

p r o t e i n a s e s ,

a n d

m e t a l l o p r o t e i n a s e s .

A t t a c h m e n t o f

a

p r o t e a s e t o

a

c e r t a i n

group

depends on

h e s t r u c t u r e

of a t a

l y t i c s i t e

a n d t h e a m i n o a c i d ( a s

o n e o f

t h e

c o n s t i t u e n t s )

e s s e n t i a l

f o r

i t s a c t i v i t y .

[ 0 0 2 5 ] P r o t e a s e s

a r e u s e d t h r o u g h o u t a n o r g a n i s m

f o r

v a r i

o u s

m e t a b o l i c p r o c e s s e s . F o r e x a m p l e ,

a c i d p r o t e a s e s

s e c r e t e d

i n t o

t h e

s t o m a c h ( s u c h a s p e p s i n ) a n d s e r i n e p r o

t e a s e s p r e s e n t

i n duodenum ( t r y p s i n

a n d

c h y m o t r y p s i n )

e n a b l e u s t o d i g e s t

t h e p r o t e i n

i n f o o d .

[ 0 0 2 6 ]

T r y p s i n i s

s e c r e t e d

i n t o

t h e d u o d e n u m , W h e r e

t a c t s

t o

h y d r o l y s e p e p t i d e s i n t o t h e i r s m a l l e r

b u i l d i n g - b l o c k s ,

namely a m i n o

a c i d s ( t h e s e

p e p t i d e s

a r e t h e

r e s u l t

o f t h e

enzyme e p s i n ’ s b r e a k i n g doWn h e

p r o t e i n s i n

t h e s t o m a c h ) .

T h i s e n a b l e s t h e

u p t a k e o f

p r o t e i n

i n

t h e

f o o d b e c a u s e p e p

J a n .

3 ,

2 0 1 3

t i d e s

( t h o u g h s m a l l e r

t h a n

p r o t e i n s ) a r e t o o

b i g

t o

b e a b s o r b e d

t h r o u g h h e l i n i n g o f h e

i l e u m .

T r y p s i n

c a t a l y s e s

t h e

h y d r o l y

s i s o f p e p t i d e b o n d s .

[ 0 0 2 7 ] The

n z y m a t i c

mechanism s

s i m i l a r

t o t h a t

o f

o t h e r

s e r i n e p r o t e a s es .

These

enzymes

o n t a i n

a

c a t a l y t i c t r i a d

con

s i s t i n g o f

h i s t i d i n e - 5 7 , a s p a r t a t e - 1 0 2 , a n d s e r i n e - 1 9 5 . T h e s e

t h r e e r e s i d u e s form

a

c h a r g e r e l a y t h a t s e r v e s t o make t h e

a c t i v e

s i t e

s e r i n e

n u c l e o p h i l i c .

T h i s

i s a c h i e v e d b y

m o d i f y i n g

t h e e l e c t r o s t a t i c

e n v i r o n m e n t o f

t h e s e r i n e . The

e n z y m a t i c

r e a c t i o n t h a t t r y p s i n s c a t a l y z e i s t h e r m o d y n a m i c a l l y f a v o r

a b l e b u t r e q u i r e s s i g n i ? c a n t a c t i v a t i o n e n e r g y ( i t

i s

“ k i n e t i

c a l l y

u n f a v o r a b l e ” ) .

I n a d d i t i o n ,

t r y p s i n c o n t a i n s a n “ o x y a n

i o n

h o l e ” f o r m e d

by

t h e b a c k b o n e a m i d e h y d r o g e n a t o m s o f

G l y - 1 9 3 a n d

S e r - 1 9 5 ,

Which s e r v e s

t o s t a b i l i z e

t h e d e v e l o p

i n g n e g a t i v e c h a r g e o n t h e c a r b o n y l o x y g e n

a t o m

o f

t h e

c l e a v e d a m i d e s .

[ 0 0 2 8 ] The s p a r t a t e r e s i d u e ( A s p

1 8 9 )

l o c a t e d i n t h e

c a t a

l y t i c p o c k e t

( S i )

o f

t r y p s i n s i s r e s p o n s i b l e f o r a t t r a c t i n g a n d

s t a b i l i z i n g

p o s i t i v e l y - c h a r g e d l y s i n e

a n d / o r a r g i n i n e ,

a n d

i s ,

t h u s ,

r e s p o n s i b l e

f o r

t h e s p e c i ? c i t y

o f

t h e

e n z y m e .

T h i s

means h a t t r y p s i n p r e d o m i n a n t l y

c l e a v e s

p r o t e i n s a t t h e c a r

b o x y l s i d e

( o r “ C - t e r m i n a l

s i d e ” ) o f t h e

a m i n o

a c i d s

l y s i n e

and

a r g i n i n e , e x c e p t When e i t h e r i s bound t o a c - t e r m i n a l

p r o l i n e .

T r y p s i n s

a r e

c o n s i d e r e d

e n d o p e p t i d a s e s , i . e . , t h e

c l e a v a g e o c c u r s W i t h i n

t h e

p o l y p e p t i d e c h a i n r a t h e r

t h a n

a t

t h e

t e r m i n a l

amino a c i d s

l o c a t e d a t

t h e

e n d s o f p o l y p e p t i d e s .

[ 0 0 2 9 ] Green e t a l . , The e f f e c t s o f d i v a l e n t c a t i o n s on

t r y p s i n ,

J . B i o l . C h e m . , 2 0 4 : 3 7 9 - 3 9 0 ( 1 9 5 3 ) , d i s c l o s e s

t h a t

m e t a l

i o n s

s u c h a s Hg++,

Cu++,

A g + and o

a l e s s e r e x t e n t

Zn++

i n h i b i t e d

t h e

e s t e r a s e

a c t i v i t y o f t r y p s i n a t loW concen

t r a t i o n s ,

i . e . ,

a r o u n d 0 . 0 0 1 M.

[ 0 0 3 0 ] T o y o t a e t

a l . , S y n t h e s i s a n d e v a l u a t i o n

o f

g u a n i d i n e

c o n t a i n i n g S c h i f f b a s e

c o p p e r

( I I ) , z i n c ( I I ) a n d i r o n

( I I )

c h e l a t e s

a s t r y p s i n i n h i b i t o r s , Chem. P h a r m . B u l l . , 5 1 ( 6 ) :

6 2 5 - 6 2 9

( 2 0 0 3 ) , d i s c l o s e s t h e s t u d y o f

S c h i f f

b a s e m e t a l

c h e l a t e s i n h i b i t o r y

a c t i v i t y a g a i n s t t r y p s i n .

[ 0 0 3 1 ] J a n e e t a l . ,

A

o v e l a p p r o a c h t o

s e r i n e p r o t e a s e

i n h i b i t i o n :

k i n e t i c

c h a r a c t e r i z a t i o n

o f

i n h i b i t o r s Who e

p o t e n

c i e s a n d

s e l e c t i v i t i e s

a r e d r a m a t i c a l l y e n h a n c e d

by

z i n c ( I I ) ,

B i o c h e m i s t r y ,

3 9 :

4 7 9 2 - 4 8 0 0

( 2 0 0 1 ) , d i s c l o s e s

t h a t

z i n c

( I I )

p o t e n t i a t e s t h e i n h i b i t o r y i n t e r a c t i o n b e t W e e n

l o W - m o l e c u l a r

W e i g h t

c h e l a t i n g

i n h i b i t o r s a n d t r y p s i n .

[ 0 0 3 2 ] P r o t e a s e s i n

t h e s k i n a r e e s s e n t i a l t o e p i d e r m a l p e r

m e a b i l i t y b a r r i e r h o m e o s t a s i s .

I n

a d d i t i o n

t o

t h e i r

d i r e c t p r o

t e o l y t i c e f f e c t s , c e r t a i n p r o t e a s e s s i g n a l

t o c e l l s

b y a c t i v a t i n g

p r o t e a s e - a c t i v a t e d r e c e p t o r s

( P A R s ) , t h e G - p r o t e i n - c o u p l e d

r e c e p t o r s .

The x p r e s s i o n o f u n c t i o n a l

PAR-2

on

human k i n

a n d i t s

r o l e

i n i n ? a m m a t i o n ,

p r u r ‘ i t u s ,

a n d s k i n

b a r r i e r

homeo

s t a s i s

h a v e

b e e n

d e m o n s t r a t e d .

A t o p i c

d e r m a t i t i s

(AD) i s a

m u l t i f a c t o r i a l i n ? a m m a t o r y

s k i n

d i s e a s e

c h a r a c t e r i z e d

b y

g e n e t i c b a r r i e r d e f e c t s a n d a l l e r g i c

i n ? a m m a t i o n , W h i c h

i s

s u s t a i n e d b y

g e n e - e n v i r o n m e n t a l

i n t e r a c t i o n s . R e c e n t s t u d i e s

h a v e r e v e a l e d a b e r r a n t e x p r e s s i o n

a n d a c t i v a t i o n

o f

s e r i n e

p r o t e a s e s a n d PAR-2 i n t h e l e s i o n a l s k i n o f AD

a t i e n t s . The

i m b a l a n c e b e t W e e n p r o t e a s e s a n d p r o t e a s e i n h i b i t o r s a s s o c i

a t e d

W i t h

g e n e t i c

d e f e c t s

i n

t h e

p r o t e a s e / p r o t e a s e

i n h i b i t o r

e n c o d i n g g e n e s , i n c r e a s e

i n

s k i n s u r f a c e

p H ,

and x p o s u r e t o

p r o t e o l y t i c a l ly a c t iv e a l l e r g e n s c o n t r i b u t e t o t h i s a b e r r a n t

p r o t e a s e / P A R - 2

s i g n a l i n g

i n AD. T h e i n c r e a s e d p r o t e a se

a c t i v i t y i n AD e a d s t o

a b n o r m a l

d e s q u a m a t i o n , d e g r a d a t i o n

o f

l i p i d - p r o c e s s i n g

e n z y m e s a n d a n t i m i c r o b i a l p e p t i d e s , a n d

a c t i v a t i o n

o f

p r i m a r y c y t o k i n e s ,

t h e r e b y l e a d i n g

t o p e r m e

a b i l i t y b a r r i e r

d y s f u n c t i o n ,

i n ? a m m a t i o n , a n d

d e f e c t s

i n t h e

a n t i m i c r o b i a l b a r r i e r .

M o r e o v e r ,

u p - r e g u l a t e d p r o t e a s e s

8/16/2019 Us 20130004580

http://slidepdf.com/reader/full/us-20130004580 7/16

US 2013/0004580

A 1

s t i m u l a t e

PAR-2 i n

l e s i o n a l s k i n

of AD and l e a d t o

t h e p r o

d u c t i o n o f

c y t o k i n e s

a n d c h e m o k i n e s

i n v o l v e d

i n in?amma

t i o n a n d immune e s p o n s e s , i t c h i n g s e n s a t i o n , a n d s u s t a i n e d

e p i d e r m a l b a r r i e r

p e r t u r b a t i o n W i t h e a s i e r a l l e r g e n p e n e t r a

t i o n .

I n a d d i t i o n ,

PAR-2 s

an

m p o r t a n t

s e n s o r

f o r

exogenous

d a n g e r

m o l e c u l e s ,

s u c h as

e x o g e n o u s p r o t e a s e s

f r o m

v a r i o u s

a l l e r g e n s , a n d p l a y s a n i m p o r t a n t r o l e i n AD a t h o g e n e s i s .

Lee et a l . d i s c l o s e t h a t p r o t e a s e a c t i v i t y o r PAR-2 m a y be a

f u t u r e t a r g e t f o r t h e r a p e u t i c i n t e r v e n t i o n f o r t h e

t r e a t m e n t

o f

AD.

S e e

Lee t

a l . ,

P r o t e a s e a n d P r o t e a s e - A c t i v a t e d Recep

t o r - 2

S i g n a l i n g

i n t h e P a t h o g e n e s i s o f A t o p i c

D e r m a t i t i s ,

Y o n s e i

Med . 2010 November ;

5 1 ( 6 ) :

8 0 8 - 8 2 2 .

[ 0 0 3 3 ] What s n e e d e d , t h e r e f o r e , i s

a

s k i n

c a r e c o m p o s i

t i o n t h a t c a n b e a p p l i e d

d i r e c t l y t o

t h e s k i n t o

b l o c k

t h e

a c t i v i t y o f e n Z y m e s s u c h a s

t r y p s i n

f o u n d i n

b o d i l y ? u i d s ,

f e c e s o r in?amed k i n . T h i s n e e d i s p r o v i d e d by t h e p r e s e n t

i n v e n t i o n .

The i n v e n t i o n

h a s p a r t i c u l a r r e l e v a n c e

t o d i a p e r

r a s h a n d a t o p i c d e r m a t i t i s .

SUMMARY

OF

THE

INVENTION

[ 0 0 3 4 ] T h e p r e s e n t i n v e n t i o n d e s c r i b e s

c o m p o s i t i o n s

a n d

m e t h o d s

f o r t r e a t i n g

a n d / o r p r e v e n t i n g i n ? a m m a t o r y

d e r m a

t o s e s ,

a n d i n p a r t i c u l a r e n z y m a t i c d e r m a t i t i s , s u c h

a s

d i a p e r

r a s h

a n d a t o p i c

d e r m a t i t i s ,

v i a t h e t o p i c a l a d m i n i s t r a t i o n o f

d i v a l e n t c a t i o n s .

[ 0 0 3 5 ]

T h e

p r e s e n t

i n v e n t i o n a l s o

d e s c r i b e s

c o m p o s i t i o n s

a n d

m e t h o d s

f o r t r e a t i n g a n d /

o r p r e v e n t i n g i n ? a m m a t o r y

d e r

m a t o s e s , a n d i n p a r t i c u l a r e n Z y m a t i c d e r m a t i t i s , s u c h as d i a

p e r

r a s h

a n d

a t o p i c d e r m a t i t i s ,

v i a t h e

t o p i c a l

a d m i n i s t r a t i o n

of i v a l e n t

c a t i o n / a n i o n p a i r s , o t h e r W i s e r e f e r r e d t o

h e r e i n

a s

d i v a l e n t c a t i o n s a l t s .

[ 0 0 3 6 ]

T h e

p r e s e n t

i n v e n t i o n a l s o

d e s c r i b e s

c o m p o s i t i o n s

a n d m e t h o d s

f o r t r e a t i n g

a n d / o r p r e v e n t i n g i n ? a m m a t o r y d e r

m a t o s e s , a n d i n p a r t i c u l a r e n Z y m a t i c d e r m a t i t i s , s u c h as d i a

p e r

r a s h

a n d

a t o p i c

d e r m a t i t i s ,

v i a t h e

t o p i c a l

a d m i n i s t r a t i o n

o f

a s p e c i ? e d d i v a l e n t

c a t i o n , i . e . ,

m a g n e s iu m , a n d

t a l c .

The

magnesium may e p r e s e n t i n

t h e c o m p o s i t i o n

i n

i t s d i v a l e n t

c a t i o n s a l t f o r m .

[ 0 0 3 7 ] T h e p r e s e n t i n v e n t i o n a l s o d e s c r i b e s c o m p o s i t i o n s

a n d

m e t h o d s

f o r t r e a t i n g a n d /

o r p r e v e n t i n g i n ? a m m a t o r y

d e r

m a t o s e s , a n d i n p a r t i c u l a r e n Z y m a t i c d e r m a t i t i s , s u c h as d i a

p e r r a s h

a n d

a t o p i c d e r m a t i t i s , v i a t h e t o p i c a l

a d m i n i s t r a t i o n

of

Z i n c

oxide

and a c i d s o a s t o r e l e a s e a speci?ed d i v a l e n t

c a t i o n ,

i . e . ,

Z i n c .

[ 0 0 3 8 ] O t h e r f e a t u r e s a n d a d v a n t a g e s o f

t h e

p r e s e n t i n v e n

t i o n W i l l b e a p p a r e n t f r o m t h e d e t a i l e d d e s c r i p t i o n o f t h e

i n v e n t i o n and from h e c l a i m s .

BRIEF

DESCRIPTION

OFTHE

FIGURES

[ 0 0 3 9 ] F I G . 1 i s a g r a p h s h o W i n g t h e

e f f e c t

o f v a r i o u s

c a t i o n s on h e i n h i b i t i o n o f t r y p s i n .

[ 0 0 4 0 ]

F I G . 2

i s a g r a p h s h o W i n g t h e

e f f e c t

o f v a r i o u s s a l t

f o r m s o f d i v a l e n t c a t i o n s on

h e

i n h i b i t i o n o f

t r y p s i n

a t v a r i

ous

c o n c e n t r a t i o n s i n mM.

[ 0 0 4 1 ]

F I G . 3 i s a g r a p h s h o W i n g

( 1 )

t h e e f f e c t o f

v a r i o u s

amounts of i t r i c

a c i d

i n

a Z i n c o x i d e c o n t a i n in g f o r m u l a t i o n

on h e i n h i b i t i o n o f t r y p s i n and 2 ) a c o m p a r i s o n o f t h e

e f f e c t

of i t r i c a c i d i n a Z i n c

o x i d e

c o n t a i n i n g

f o r m u l a t i o n

on t h e

i n h i b i t i o n

o f

t r y p s i n

W i t h

t h e

e f f e c t

o f

Baby

P a s t e 5 . 0 , a

c o m m e r c i a l l y a v a i l a b l e

p r o d u c t

f r o m M e n d e l s o n P h a r m a

c e u t i c a l s LLC, M o u n t a i n s i d e , N . J . , on t h e

i n h i b i t i o n

o f

t r y p s i n .

J a n .

3 ,

2 0 1 3

DETAILED DESCRIPTION OF THE INVENTION

[ 0 0 4 2 ]

U n l e s s d e ? n e d

o t h e r W i s e ,

a l l t e c h n i c a l a n d s c i e n

t i ? c t e r m s

used

e r e i n

have

t h e same meaning

a s

commonly

u n d e r s t o o d

by one o f o r d i n a r y

s k i l l

i n t h e a r t

t o

Which

t h e

i n v e n t i o n b e l o n g s . A l s o ,

a l l

p u b l i c a t i o n s ,

p a t e n t

a p p l i c a t i o n s ,

p a t e n t s , and o t h e r r e f e r e n c e s mentioned

e r e i n a r e i n c o r p o

r a t e d

by

r e f e r e n c e . As

u s e d

h e r e i n , a l l p e r c e n t a g e s a r e b y

W e i g h t u n l e s s

o t h e r W i s e

s p e c i ? e d .

[ 0 0 4 3 ] As s e d

h e r e i n , “ t r y p s i n

i n h i b i t o r y

a c t i v i t y ”

m e a n s

t h e a b i l i t y t o i n h i b i t t h e a c t i v i t y

o f

t h e p r o t e a s e , t r y p s i n ,

as

measured

by h e

a s s a y s e t

f o r t h

beloW n

t h e

e x a m p l e s . I n o n e

e m b o d i m e n t ,

t h e

d i v a l e n t

c a t i o n s u s e d

i n

t h e p r e s e n t i n v e n

t i o n

h a v e a t r y p s i n i n h i b i t o r y

a c t i v i t y

o f t

l e a s t a b o u t

15 . n

a

f u r t h e r embodiment,

h e

d i v a l e n t

c a t i o n s u s e d i n t h e

p r e s e n t

i n v e n t i o n h a v e a

t r y p s i n i n h i b i t o r y

a c t i v i t y

o f a t

l e a s t a b o u t

25 , and r e f e r a b l y a t l e a s t a b o u t 50 , r a t l e a s t a b o u t 60 ,

o r at l e a s t about 70 , o r a t l e a s t about

80 ,

o r a t l e a s t about

90 .

[ 0 0 4 4 ]

As

s e d h e r e i n ,

“ p r e v e n t ” o r “ p r e v e n t i n g ” means

o

p r o a c t i v e l y

s t o p

t h e

d e v e l o p m e n t

o f e n Z y m a t i c

s k i n

i r r i t a t i o n ,

o r t o h a l t o r sloW doWn h e

p r o g r e s s i o n

of

such

r r i t a t i o n , o r t o

r e d u c e

t h e

r i s k

o f d e v e l o p i n g

s u c h i r r i t a t i o n .

[ 0 0 4 5 ] As

u s e d

h e r e i n , “ t r e a t ” o r “ t r e a t i n g ”

i s meant t o

comfort h e

s k i n

n e a r a n d / o r a t t h e l o c a t i o n o f enZymatic

s k i n

i r r i t a t i o n , a n d i f p o s s i b l e to i n d u c e a r e g r e s s i o n i n s u c h

i r r i

t a t i o n .

A s

u s e d

h e r e i n , “ r e g r e s s i o n ”

i s

meant

o r e d u c e

t h e

a m o u n t

a n d / o r

s e v e r i t y

o f

t o p i c a l

e n z y m a t i c

s k i n i r r i t a t i o n

s y m p t o m s ,

s u c h

a s , e . g . , i r r i t a t i o n , r e d n e s s ,

b l i s t e r s , d i s c o m

f o r t , e x c o r i a t i o n ,

a n d

t h e l i k e .

[ 0 0 4 6 ]

As

s e d e r e i n , “ t o p i c a l a p p l i c a t i o n ”

m e a n s

d i r e c t l y

l a y i n g on r s p r e a d i n g

on

u t e r s k i n

u s i n g ,

e . g . , t h e

h a n d s ,

an

a p p l i c a t o r such

a s

a W i p e ,

p u f f , r o l l e r , o r

s p r a y ,

o r

v i a a

s u b s t r a t e such a s a d i a p e r .

[ 0 0 4 7 ]

As

used e r e i n , “ a f f e c t e d a r e a ”

i s

meant h e a r e a

o f

s k i n

t h a t

i s

p r e s e n t l y e x h i b i t i n g

a n y

l e v e l

o f

s k i n r a s h o r

e n Z y m a t i c

d e r m a t i t i s ,

o r

t h e a r e a

t h a t

W i l l

b e i n p r o l o n g e d

c o n t a c t

W i t h

u r i n e o r f e c e s c o n t a i n i n g

s u c h

d e r m a t i t i s - c a u s

i n g enZymes. T h i s a r e a a l s o i n c l u d e s t h e a r e a

i m m e d i a t e l y

proximate t o t h e d e s c r i b e d a r e a .

I t

i s t h e a r e a a t

Which

r e a t

ment a n d / o r p r e v e n t i o n

i s

d e s i r e d .

[ 0 0 4 8 ] A s used e r e i n , “ t o p i c a l

c a r r i e r ”

means one r more

c o m p a t i b l e s o l i d

o r

l i q u i d ? l l e r d i l u e n t s t h a t

a r e

s u i t a b l e f o r

t o p i c a l

a d m i n i s t r a t i o n t o a

mammal.

E x a m p l e s o f

t o p i c a l

c a r r i e r s i n c l u d e , b u t a r e n o t l i m i t e d

t o ,

W a t e r ,

W a x e s , o i l s ,

e m o l l i e n t s , e m u l s i ? e r s , t h i c k e n i n g a g e n t s , g e l l i n g

a g e n t s , a n d

m i x t u r e s t h e r e o f .

[ 0 0 4 9 ]

T h e

t o p i c a l c o m p o s i t i o n s o f t h e p r e s e n t i n v e n t i o n

comprise a s a f e and f f e c t i v e amount of d i v a l e n t c a t i o n . I n

o n e

e m b o d i m e n t , t h e c o m p o s i t i o n c o n t a i n s , b a s e d

upon

t h e

t o t a l

amount o f t o p i c a l c o m p o s i t i o n ,

from a b o u t

0.001 o

a b o u t 50% i v a l e n t

c a t i o n .

I n a n o t h e r embodiment, t h e com

p o s i t i o n c o n t a i n s from

a b o u t 0.001

to

a b o u t 3 0 d i v a l e n t

c a t i o n . I n a p r e f e r r e d

e m b o d i m e n t ,

t h e

c o m p o s i t i o n

c o n t a i n s

from about

0 . 0 1 t o about

10% d i v a l e n t c a t i o n . I n a more

p r e f e r r e d e m b o d i m e n t , t he

c o m p o s i t i o n

c o n t a i n s

f r o m

a b o u t

0 . 1

t o

a b o u t

5.0

d i v a l e n t

c a t i o n . I n a most p r e f e r r e d

e m b o d i m e n t , t h e c o m p o s i t i o n

c o n t a i n s

f r o m

a b o u t 0.1 o

a b o u t 1.0 d i v a l e n t c a t i o n .

[ 0 0 5 0 ] I n

o n e

e m b o d i m e n t , t h e c o m p o s i t i o n c o n t a i n s ,

b a s e d

upon

t h e t o t a l

amount

o f t o p i c a l c o m p o s i t i o n , f r o m

a b o u t

0.001

to a b o u t 5 0 d i v a l e n t c a t i o n s a l t . I n a n o t h e r

e m b o d i m e n t ,

t h e

c o m p o s i t i o n

c o n t a i n s f r o m a b o u t 0.001 o

a b o u t

3 0 d i v a l e n t c a t i o n s a l t . I n a p r e f e r r e d e m b o d i m e n t ,

t h e c o m p o s i t i o n

c o n t a i n s

from a b o u t 0.01 to a b o u t 1 0

8/16/2019 Us 20130004580

http://slidepdf.com/reader/full/us-20130004580 8/16

US 2013/0004580

A 1

d i v a l e n t c a t i o n s a l t . I n a

more

p r e f e r r e d embodiment, t h e

c o m p o s i t i o n c o n t a i n s from

b o u t 0 .

1 o

a b o u t

5.0 i v a l e n t

c a t i o n

s a l t .

I n a most

r e f e r r e d e m b o d i m e n t ,

t h e

c o m p o s i t i o n

c o n t a i n s

from

a b o u t

0.1 o

a b o u t 1.0

d i v a l e n t

c a t i o n

s a l t .

[ 0 0 5 1 ] I n

o n e e m b o d i m e n t ,

t h e

c o m p o s i t i o n

c o n t a i n s ,

f r o m

a b o u t 0 . 1 mM

o

a b o u t 5000 mM i v a l e n t c a t i o n s a l t . I n

a n o t h e r e m b o d i m e n t ,

t h e

c o m p o s i t i o n

c o n t a i n s f r o m a b o u t

0 . 1

mM o

a b o u t

3000 mM

i v a l e n t c a t i o n s a l t . I n

a

p r e f e r r e d

e m b o d i m e n t ,

t h e

c o m p o s i t i o n c o n t a i n s

from

b o u t 1 . 0 mM

o

a b o u t

1000

mM

i v a l e n t

c a t i o n s a l t .

I n

a more p r e f e r r e d

embodiment h e c o m p o s i t i o n

c o n t a i n s

from b o u t 1 . 0 mM o

a b o u t 500 mM i v a l e n t

c a t i o n

s a l t . I n a most p r e f e r r e d

e m b o d i m e n t ,

t h e

c o m p o s i t i o n

c o n t a i n s from b o u t

1 . 0 mM

o

a b o u t 500

mM

i v a l e n t c a t i o n s a l t .

[ 0 0 5 2 ] I n a n e m b o d i m e n t , t h e p r e s e n t i n v e n t i o n

d i s c l o s e s

t h e u s e

o f

d i v a l e n t c a t i o n s i n c o m p o s i t i o n s f o r t h e

t r e a t m e n t

o f

d i a p e r d e r m a t i t i s ( d i a p e r r a s h ) .

[ 0 0 5 3 ]

The

c o m p o s i t i o n s

u s e f u l i n t h e

p r e s e n t i n v e n t i o n

i n v o l v e

f o r m u l a t i o n s s u i t a b l e f o r t o p i c a l a p p l i c a t i o n t o

s k i n .

I n

o n e e m b o d i m e n t ,

t h e

c o m p o s i t i o n

c o n t a i n s

a n

a c c e p t a b l e

t o p i c a l

c a r r i e r

i n a n a m o u n t , b a s e d upon h e

t o t a l

W e i g h t o f t h e

c o m p o s i t i o n ,

f r o m

a b o u t

5 0

o a b o u t 9 9 . 9 9 % , e . g . , from

a b o u t

80%

o

a b o u t

9 5 .

[ 0 0 5 4 ]

The

o m p o s i t i o n may e made n t o a W i d e v a r i e t y o f

p r o d u c t

t y p e s

t h a t

i n c l u d e b u t a r e n o t l i m i t e d

t o l o t i o n s ,

c r e a m s , g e l s ,

s t i c k s , s p r a y s ,

o i n t m e n t s ,

c l e a n s i n g l i q u i d

W a s h e s a n d s o l i d b a r s , p a s t e s , p o W d e r s , m o u s s e s , W i p e s ,

p a t c h e s , Wound

r e s s i n g

a n d

a d h e s i v e

b a n d a g e s , h y d r o g e l s ,

a n d ? l m s . T h e s e p r o d u c t t y p e s may o m p r i s e s e v e r a l t y p e s o f

a c c e p t a b l e t o p i c l c a r r i e r s i n c l u d i n g , b u t n o t l i m i t e d t o s o l u

t i o n s ,

e m u l s i o n s

( e . g . , m i c r o e m u l s i o n s a n d n a n o e m u l s i o n s ) ,

g e l s , s o l i d s

a n d l i p o s o m e s . The o l l o W i n g a r e n o n - l i m i t a t i v e

e x a m p l e s o f s u c h c a r r i e r s . O t h e r

c a r r i e r s

c a n be f o r m u l a t e d

by

t h o s e

o f

o r d i n a r y s k i l l

i n t h e a r t .

[ 0 0 5 5 ]

T h e

t o p i c a l c o m p o s i t i o n s u s e f u l i n t h e p r e s e n t

i n v e n t i o n c a n be o r m u l a t e d a s

anhydrous r o d u c t s .

As used

h e r e i n , “ a n h y d r o u s ” s h a l l mean h a t

t h e

c o m p o s i t i o n s c o n t a i n

l e s s

t h a n a b o u t

10 , e . g . , l e s s

t h a n

a b o u t 5%

r

l e s s

t h a n

about

1%

a t e r .

[ 0 0 5 6 ]

A l t e r n a t i v e l y , t h e t o p i c a l c o m p o s i t i o n s u s e f u l

i n

t h e

p r e s e n t i n v e n t i o n

can be

formulated a s s o l u t i o n s . S o l u t i o n s

t y p i c a l l y i n c l u d e W a t e r ( e . g . ,

f r o m

a b o u t 5 0 t o a b o u t

99.99 o r from a b o u t 90% o a b o u t 99% f W a t e r ) .

[ 0 0 5 7 ] T o p i c a l

c o m p o s i t i o n s u s e f u l

i n

t h e s u b j e c t i n v e n t i o n

may be

formulated

a s a s o l u t i o n comprising an

e m o l l i e n t .

Such o m p o s i t i o n s may o n t a i n from b o u t 2%

o

a b o u t 5 0

o f a n

e m o l l i e n t ( s ) .

A s

used e r e i n , “ e m o l l i e n t s ” r e f e r

t o

mate

r i a l s

used

o r t h e p r e v e n t i o n o r r e l i e f of r y n e s s , a s W e l l a s f o r

t h e p r o t e c t io n o f t h e

s k i n .

A i d e v a r i e t y o f

s u i t a b l e

e m o l

l i e n t s a r e

knoWn and

may

e u s e d

h e r e i n . S a g a r i n ,

C o s m e t i c s ,

S c i e n c e

a n d

T e c h n o l o g y ,

2 n d

E d i t i o n , V o l .

1 ,

p p .

3 2 - 4 3

( 1 9 7 2 )

a n d

t h e I n t e r n a t i o n a l C o s m e t i c I n g r e d i e n t D i c t i o n a r y

a n d H a n d b o o k , e d s . W e n n i n g e r a n d

McEWen,

p .

1 6 5 6 - 6 1 ,

1 6 2 6 , a n d

1 6 5 4 - 5 5 ( T h e C o s m e t i c , T o i l e t r y , a n d F r a g r a n c e

A s s o c . , W a s h i n g t o n , DC, . s u p . t h E d i t i o n , 1 9 9 7 ) ( h e r e i n a f

t e r

“ C I

Handbook”) c o n t a i n numerous

e x a m p l e s o f

s u i t a b l e

m a t e r i a l s .

[ 0 0 5 8 ] A o t i o n

can

be

made from such a s o l u t i o n . L o t i o n s

t y p i c a l l y

c o m p r i s e

f r o m a b o u t 1 o a b o u t 2 0

e . g . ,

f r o m

a b o u t 5%

o

a b o u t 10 ) o f

an

e m o l l i e n t ( s ) and from a b o u t

5 0 o

a b o u t

9 0 e . g . , from a b o u t 60 o

a b o u t

80 ) o f

Water.

[ 0 0 5 9 ] A n o t h e r

t y p e o f

p r o d u c t

t h a t may

b e f o r m u l a t e d

from s o l u t i o n

i s

a

cream. A

ream y p i c a l l y comprises from

J a n .

3 ,

2 0 1 3

a b o u t

5%

o a b o u t 5 0

e . g . ,

from b o u t

1 0

o a b o u t

20 )

o f a n e m o l l i e n t ( s ) and from

a b o u t

4 5

t o a b o u t

8 5

e . g . ,

from

a b o u t 5 0 o a b o u t 75 )

o f W a t e r .

[ 0 0 6 0 ] Y e t a n o t h e r ty p e

o f p r o d u c t t h a t may e f o r m u l a t e d

from

a s o l u t i o n

i s

an

i n t m e n t .

An

intment may

omprise

a

s i m p l e

b a s e

o f

a n i m a l o r

v e g e t a b l e

o i l s o r s e m i - s o l i d

h y d r o

c a r b o n s .

An

i n t m e n t

may c o m p r i s e from

b o u t

2% o

a b o u t

1 0 o f a n e m o l l i en t ( s ) p l u s from

a b o u t

0.1

o a b o u t

2% f

a

t h i c k e n i n g

a g e n t ( s ) .

A ore c o m p l e t e

d i s c l o s u r e

o f

t h i c k

e n i n g a g e n t s o r v i s c o s i t y i n c r e a s i n g a g e n t s u s e f u l h e r e i n c a n

b e f o u n d

i n

S a g a r i n , C o s m e t i c s , S c i e n c e a n d

T e c h n o l o g y ,

2 n d

E d i ti o n , V o l .

1 ,

p p . 7 2 - 7 3 ( 1 9 7 2 ) a n d t h e I C I Handbook p .

1 693

1 6 9 7 .

[ 0 0 6 1 ]

T h e

t o p i c a l

c o m p o s i t i o n s u s e f u l i n t h e p r e s e n t

i n v e n t i o n can

l s o

be formulated s e m u l s i o n s . I f t h e c a r r i e r i s

an

e m u l s i o n , from

a b o u t

1%

o a b o u t

1 0 e . g . , from

a b o u t

2% o

a b o u t 5 )

o f t h e

c a r r i e r c o m p r i s e s

a n e m u l s i ? e r ( s ) .

E m u l s i ? e r s may be n o n i o n i c , a n i o n i c o r c a t i o n i c . S u i t a b l e

e m u l s i ? e r s a r e

d i s c l o s e d

i n ,

e . g . , US. P a t .

No.

3 , 7 5 5 , 5 6 0 ,

US. P a t .

N o .

4 , 4 2 1 , 7 6 9 ,

M c C u t c h e o n ’ s

D e t e r g e n t s

a n d

E m u l s i ? e r s ,

N o r t h

A m e r i c a n E d i t i o n , p p . 3 1 7 - 3 2 4 ( 1 9 8 6 ) ,

and h e

I C I

Handbook, p . 1 6 7 3 - 1 6 8 6 .

[ 0 0 6 2 ]

Lotions and

creams

can be

formulated s

emulsions.

T y p i c a l l y s u c h l o t i o n s c o m p r i s e from 0.5 o

a b o u t

5%

f

a n

e m u l s i ? e r ( s ) .

S u c h c r e a m s W o u l d

t y p i c a l l y c o m p r i s e

f r o m

a b o u t

1 o a b o u t

2 0

e . g . , from

b o u t

5% o a b o u t 10 ) f

an

e m o l l i e n t ( s ) ;

from a b o u t 2 0 to a b o u t

8 0

( e . g . , from

3 0

o a b o u t 70 ) f a t e r ;

and from

b o u t

1

o a b o u t 1 0

( e . g . , f r o m

a b o u t

2%

o a b o u t 5 )

o f a n e m u l s i ? e r ( s ) .

[ 0 0 6 3 ] S i n g l e

e m u l s i o n

s k i n c a r e p r e p a r a t i o n s , s u c h a s

l o t i o n s

and

c r e a m s , o f t h e

o i l - i n - W a t e r

t y p e

and a t e r - i n - o i l

t y p e a r e Well-knoWn n t h e c o s m e t i c

a r t

and a r e u s e f u l i n t h e

s u b j e c t i n v e n t i o n . M u l t i p h a s e e m u l s i o n c o m p o s i t i o n s ,

s u c h

a s t h e W a t e r - i n - o i l - i n - W a t e r t y p e , a s d i s c l o s e d i n

US. P a t .

N o s . 4 , 2 5 4 , 1 0 5 a n d 4 , 9 6 0 , 7 6 4 ,

a r e

a l s o

u s e f u l i n t h e s u b j e c t

i n v e n t io n . I n g e n e r a l , s u c h s i n g l e o r m u l t i p h a s e e m u l s i o n s

c o n t a i n

W a t e r , e m o l l i e n t s , a n d e m u l s i ? e r s a s e s s e n t i a l i n g r e

d i e n t s .

[ 0 0 6 4 ] The

o p i c a l c o m p o s i t i o n s

o f h i s i n v e n t i o n c a n

a l s o

be f o r m u l a t e d a s a g e l

( e . g . , a n aqueous

g e l u s i n g a s u i t a b l e

g e l l i n g

a g e n t ( s ) ) .

S u i t a b l e g e l l i n g

a g e n t s

f o r a q u e o u s g e l s

i n c l u d e , b u t a r e n o t l i m i t e d

t o ,

n a t u r a l

g u m s , a c r y l i c

a c i d

and

a c r y l a t e p o l y m e r s

a n d

c o p o l y m e r s ,

a n d

c e l l u l o s e d e r i v a t i v e s

( e . g . , h y d r o x y m e t h y l c e l l u l o s e a n d

h y d r o x y p r o p y l c e l l u l o s e ) .

S u i t a b l e g e l l i n g

a g e n t s

f o r o i l s ( s u c h a s m i n e r a l o i l ) i n c l u d e ,

b u t

a r e

n o t l i m i t e d t o , h y d r o g e n a t e d b u t y l e n e / e t h y l e n e / s t y

r e n e

c o p o l y m e r

a n d

h y d r o g e n a t e d

e t h y l e n e / p r o p y l e n e / s t y

r e n e c o p o l y m e r .

S u c h g e l s t y p i c a l l y c o m p r i s e s b e t W e e n

a b o u t

0 . 1 % a n d

5 % , b y

W e i g h t ,

o f s u c h g e l l i n g a g e n t s .

[ 0 0 6 5 ] The

o p i c a l c o m p o s i t i o n s

o f h i s i n v e n t i o n c a n

a l s o

b e

combined

i t h

p a r t i c u l a t e s

s u c h a s

c l a y ,

s i l i c a

and

s t a r c h

s u c h a s c o r n s t a r c h , a n d t h e l i k e , a n d o p t i o n a l

?oW

g e n t s

s u c h

a s t r i c a l c i u m p h o s p h a t e , i n o r d e r

t o

f o r m

d u s t i n g

p o W d e r s .

E x a m p l e s

o f s u c h

p o W d e r

c o m p o n e n t s a n d m e t h o d s

f o r mak

i n g poWder o m p o s i t i o n s

may

e f o u n d n ,

e .

g . ,

US.

a t . N o s .

4 , 5 6 8 , 5 3 9 a n d

4 , 4 8 5 , 0 9 2 .

[ 0 0 6 6 ] L i p o s o m a l f o r m u l a t i o n s a r e a l s o u s e f u l c o m p o s i

t i o n s

o f

t h e s u b j e c t i n v e n t i o n . E x a m p l e s

o f

l i p o s o m e s a r e

u n i l a m e l l a r ,

m u l t i l a m e l l a r ,

a n d

p a u c i l a m e l l a r l i p o s o m e s ,

Which

m a y

o r

m a y

n o t c o n t a i n p h o s p h o l i p i d s .

Such

compo

s i t i o n s c a n b e p r e p a r e d

b y ? r s t c o m b i n i n g

h e s p e r e t i n

W i t h

a

p h o s p h o l i p i d ,

s u c h

a s

d i p a l m i t o y l p h o s p h a t i d y l

c h o l i n e , c h o

l e s t e r o l and a t e r a c c o r d i n g t o t h e method e s c r i b e d i n MeZei

&

u l a s e k h a r a m ,

“ L i p o s o m e s i A

S e l e c t i v e D r u g

D e l i v e r y

8/16/2019 Us 20130004580

http://slidepdf.com/reader/full/us-20130004580 9/16

US 2013/0004580

A 1

S y s t e m f o r

t h e T o p i c a l R o u t e

o f

d m i n i s t r a t i o n ;

G e l

D o s a g e

F o r m ” ,

J o u r n a l

o f P h a r m a c e u t i c s a n d P h a r m a c o l o g y , V o l . 3 4

( 1 9 8 2 ) , p p .

4 7 3 - 4 7 4 ,

o r a

m o d i ? c a t i o n t h e r e o f . E p i d e r m a l

l i p i d s o f

s u i t a b l e c o m p o s i t i o n

f o r

f o r m i n g

l i p o s o m e s may e

s u b s t i t u t e d

f o r

t h e

p h o s p h o l i p i d .

T h e

l i p o s o m e p r e p a r a t i o n

may h e n i n c o r p o r a t e d

i n t o

one

o f

t h e above

c a r r i e r s

( e . g . , a

g e l o r a n o i l - i n - W a t e r e m u l s i o n )

i n

o r d e r

t o p r o d u c e t h e

l i p o

s o m a l f o r m u l a t i o n .

O t h e r

c o m p o s i t i o n s

a n d

u s e s

o f t o p i c a l l y

a p p l i e d

l i p o s o m e s

a r e d e s c r i b e d

i n M e Z e i , M . ,

“ L i p o s o m e s

as

a S k i n D r u g D e l i v e r y S y s t e m ” , T o p i c s i n P h a r m a c e u t i c a l

S c i e n c e s

( D . D . B r e i m e r a n d

P . S p e i s e r , e d s . ) , E l s e v i e r

S c i

e n c e P u b l i s h e r s

B .

V . ,

N e W

o r k , N . Y . , 1 9 8 5 , p p .

3 4 5 - 3 5 8 ,

P CT a t e n t

A p p l i c a t i o n

No. WO96/31194 and

US.

a t . No.

5 , 2 6 0 , 0 6 5 .

[ 0 0 6 7 ]

T h e t o p i c a l

c o m p o s i t i o n s

u s e f u l i n t h e s u b j e c t

i n v e n t i o n may c o n t a i n , i n a d d i t i o n t o t h e a f o r e m e n t i o n e d

c o m p o n e n t s ,

a

Wide

a r i e t y o f a d d i t i o n a l o i l - s o l u b l e m a t e r i

a l s a n d / o r W a t e r - s o l u b l e m a t er i a l s c o n v e n t i o n a l l y u s e d

i n

compositions f o r use on s k i n a t t h e i r a r t - e s t a b l i s h e d

l e v e l s .

[ 0 0 6 8 ] I n

one embodiment,

t h e

c o m p o s i t i o n

may

l s o

con

t a i n one

more of h e f o l l o W i n g : a n t i f u n g a l s such s ketocona

Z o l e , m i c o n a Z o l e , e l u b i o l , a l l a n t o i n , c a l a m i n e ,

s i l i c o n - b a s e d

o r g a n i c p o l y m e r s

s u c h

a s , b u t n o t l i m i t ed t o , p o l y m e r i z e d

s i l o x a n e

s u c h

a s

d i m e t h i c o n e ,

k a o l i n , p e t r o l a t u m , W h i t e

p e t

r o l a t u m ,

c o d l i v e r

o i l , l a n o l i n ,

m i n e r a l

i l ,

t a l c ,

t o p i c a l

s t a r c h ,

and any

o t h e r

a g e n t s u i t a b l e f o r u s e

i n t h e

t r e a t m e n t a n d / o r

p r e v e n t i o n

o f

d i a p e r r a s h .

[ 0 0 6 9 ] I n

a n o t h e r e m b o d i m e n t , t h e c o m p o s i t i o n

may

c o n

t a i n one

o r more f

t h e f o l l o w i n g

a g e n t s

i n an

amount,

based

upon

h e t o t a l W e i g h t o f t h e

c o m p o s i t i o n ,

f r o m a b o u t

0.5

o

a b o u t 2%

f

a l l a n t o i n , from b o u t

1

o a b o u t

2 5

a l a m i n e ,

from

b o u t 1% o a b o u t 30%

i m e t h i c o n e ,

from

a b o u t 4% o

a b o u t 20% a o l i n ,

from

a b o u t 30% o l e s s t h a n a b o u t 1 0 0

p e t r o l a t u m ,

from b o u t 3 0 o

l ess

t h a n a b o u t 1 0 0 f W h i t e

p e t r o l a t u m , from b o u t

5% o

a b o u t

1 4 f

cod i v e r

o i l , e . g . ,

s u c h

t h a t t h e

amount o f cod i v e r

o i l

d o e s

n o t

e x c e e d 1 0 , 0 0 0

USP

n i t s

o f

vitaminA

and

400

USP n i t s o f

c h o l e c a l c i f e r o l ,

a b o u t 1 0 o a b o u t

1 6 o f

l a n o l i n , from a b o u t 50% o l e s s

t h a n a b o u t 1 0 0 m i n e r a l

o i l ,

and from a b o u t 1 0 t o a b o u t

8 7 o p i c a l

s t a r c h .

Examples f

i t a m i n s

i n c l u d e ,

b u t

a r e

n o t

l i m i t e d t o , v i t a m i n A , v i t a m i n

Bs

s u c h

a s

v i t a m i n B 3 , v i t a m i n

B 5 , a n d v i t a m i n

B 1 2 ,

v i t a m i n C , v i t a m i n K , a n d v i t a m i n

E

and d e r i v a t i v e s t h e r e o f .

[ 0 0 7 0 ] The r e s e n t i n v e n t i o n d i s c l o s e s a c o m p o s i t i o n com

p r i s i n g ; ( i )

a d i v a l e n t

c a t i o n ,

W h e r e i n

( i i )

s a i d

c o m p o s i t i o n i s

f o r t h e

t r e a t m e n t

of

o n d i t i o n s s u c h

a s

d i a p e r d e r m a t i t i s . S a i d

composition

can

i n c l u d e

an

a c c e p t a b l e c a r r i e r o r

b a s e .

The

d i v a l e n t

c a t i o n

i s s e l e c t e d f r o m , b u t n o t l i m i t e d t o , e . g . , c o p

p e r

( 1 1 ) , Z i n c ,

i r o n

( 1 1 ) ,

c a l c i u m , s t r o n t i u m , m a g n e s i u m ,

t i n

( H ) ,

n i c k e l

( 1 1 ) ,

m a n g a n e s e

( 1 1 ) ,

a n d c a d m i u m

( 1 1 ) .

[ 0 0 7 1 ]

The

r e s e n t i n v e n t i o n

d i s c l o s e s a c o m p o s i t i o n com

p r i s i n g ;

( i )

a

d i v a l e n t c a t i o n / a n i o n p a i r ( o t h e r W i s e knoWn

s a

d i v a l e n t c a t i o n

s a l t ) ,

W h e r e i n ( i i )

sai d c o m p o s i t i o n i s

f o r t h e

t r e a t m e n t of o n d i t i o n s

such

a s d i a p e r d e r m a t i t i s . S a i d com

p o s i t i o n can

i n c l u d e

an

a c c e p t a b l e c a r r i e r o r b a s e . The

d i v a

l e n t

c a t i o n / a n i o n p a i r i s s e l e c t e d

f r o m , b u t i s n o t

l i m i t e d t o ,

t h e

d i v a l e n t

c ati o n s l i s t e d a b o v e ,

i . e . ,

c o p p e r ( 1 1 ) , Z i n c , i r o n

( 1 1 ) , c a l c i u m , s t r o n t i u m , m a g n e s i u m , t i n ( l l ) , n i c k e l ( 1 1 ) , man

g a n e s e ( I I ) , a n d cadmium H ) , i n c o m b i n a t i o n W i t h a n i o n s

s e l e c t e d f r o m ,

e . g . , v a r i o u s

c a r b o x y l i c

a c i d s

s e l e c t e d f r o m ,

b u t n o t

l i m i t e d t o ,

a c e t i c

a c i d , a s c o r b i c a c i d , a s p a r t i c a c i d ,

b e n Z o i c a c i d , b o r i c

a c i d ,

c a r b o n i c

a c i d ,

c i t r i c a c i d , f o l i c

a c i d ,

g l u c o n i c a c i d , g l u c u r o n i c a c i d , g l u t a m i c a c i d ,

g l y c y r r h i Z i c

a c i d ,

g l y o x y l i c a c i d , h y d r o c h l o r i c

a c i d ,

h y a l u r o n i c

a c i d , l a c

J a n .

3 ,

2 0 1 3

t i c a c i d ,

l a n o l i c a c i d , m a l i c a c i d , n i a c i n ,

p o l y a c r y l i c a c i d ,

r o s a m a r i n i c a c i d ,

s a l i c y l i c

a c i d ,

s o r b i c a c i d a n d

t a r t a r i c a c i d .

Examples o f

d i v a l e n t

c a t i o n / a n i o n p a i r s

i n c l u d e ,

b u t

a r e

n o t

l i m i t e d

t o , e . g . , c o p p e r ( l l ) c h l o r i d e , c o p p e r

( l l ) a c e t a t e , c o p

p e r

( 1 1 )

g l u c o n a t e ,

c o p p e r

( l l )

l a c t a t e ,

Z i n c

c h l o r i d e ,

Z i n c

a c e t a t e ,

Z i n c g l u c o n a t e , Z i n c l a c t a t e , Z i n c c a r b o n a t e , Z i n c

b o r a t e , Z i n c a s p a r t a t e , Z i n c p y r r o l i d o n e c a r b o x y l i c

a c i d ,

Z i n c

c i t r a t e , i r o n ( 1 1 ) c h l o r i d e , i r o n ( 1 1 )

a c e t a t e ,

i r o n ( 1 1 )

g l u c o n a t e ,

i r o n ( 1 1 ) l a c t a t e , c a l c i u m c h l o r i d e , c a l c i u m

a c e t a t e ,

c a l c i u m

g l u c o n a t e , c a l c i u m l a c t a t e ,

s t r o n t i u m

c h l o r i d e ,

s t r o n t i u m

a c e t a t e ,

s t r o n t i u m g l u c o n a t e , s t r o n t i u m l a c t a t e , m a g n e s i u m

c h l o r i d e ,

m a g n e s i u m

a c e t a t e ,

m a g n e s i u m

g l u c o n a t e a n d

m a g n e s i u m l a c t a t e .

[ 0 0 7 2 ]

The

r e s e n t i n v e n t i o n d i s c l o s e s

a c o m p o s i t i o n com

p r i s i n g ; ( i ) a

s p e c i ? e d

d i v a l e n t

c a t i o n ,

i . e . , m a g n e s i u m , i n

c o m b i n a t i o n

W i t h

a l c , W h e r e i n i i )

s a i d

c o m p o s i t i o n

i s

f o r t h e

t r e a t m e n t of o n d i t i o n s

such

a s d i a p e r d e r m a t i t i s . S a i d com

p o s i t i o n

c a n i n c l u d e a n

a c c e p t a b l e c a r r i e r o r

b a s e . According

t o

a n embodiment,

t h e

c o m p o s i t i o n

may

o n t a i n

from

a b o u t

45%

o l e s s

t h a n a b o u t

1 0 0 a l c .

[ 0 0 7 3 ] The r e s e n t i n v e n t i o n d i s c l o s e s a c o m p o s i t i o n

com

p r i s i n g ; ( i )

Z i n c

o x i d e

a n d a n a c i d ,

t h a t When combined

r e l e a s e s a

s p e c i ? e d

d i v a l e n t

c a t i o n ,

i . e . , Z i n c , W h e r e i n i i ) s a i d

c o m p o s i t i o n i s

f o r t h e

t r e a t m e n t o f c o n d i t i o n s s u c h a s d i a p e r

d e r m a t i t i s .

Any

a c i d t h a t

Would e s u l t

in t h e r e l e a s e o f Z i n c

c a t i o n

When

e a c t e d W i t h

Z i n c

o x i d e

can

be

s e d .

Such

c i d s

i n c l u d e ,

b u t a r e

n o t

l i m i t e d t o , e . g . , c i t r i c

a c i d ,

a c e t i c

a c i d ,

s a l i c y c l i c

a c i d ,

c a r b o n i c

a c i d ,

c l u c u r o n i c a c i d , g l u c o n i c a c i d ,

l a c t i c

a c i d

a n d

m a l i c a c i d .

P r e f e r a b l y , Z i n c

o x i d e i s p r e s e n t

i n

an amount

s e l e c t e d from

t h e

group

c o n s i s t i n g

of from

a b o u t

1% o a b o u t 40 , from a b o u t 1% o a b o u t 25 , from a b o u t

1% o a b o u t 20 , from a b o u t 1% o a b o u t 15 , from a b o u t

5% o a b o u t

13 ,

a b o u t 1 . 0 % , a b o u t

1 0 % ,

a n d a b o u t

13 .

P r e f e r a b l y

t h e a c i d

i s

p r e s e n t

i n

an

amount

s e l e c t e d

from

h e

g r o u p c o n s i s t i n g o f from

a b o u t 0.001 o a b o u t

10 , from

a b o u t 0.01 to a b o u t 5 , from a b o u t

0. 05

t o a b o u t

2.5

and

from

a b o u t 0.1

t o

a b o u t 1 . 0 % . The c o m p o s i t i o n

can

i n c l u d e an a c c e p t a b l e c a r r i e r o r b a s e .

[ 0 0 7 4 ]

A c c e p t a b l e t o p i c a l c a r r i e r s a r e

s e l e c t e d

f r o m t r a d i

t i o n a l W a t e r a n d o i l e m u l s i o n s , s u s p e n s i o n s , c o l l o i d s , m i c r o

e m u l s i o n s ,

c l e a r s o l u t i o n s ,

s u s p e n s i o n s

o f n a n o p a r t i c l e s ,

e m u l s i o n s

o f

n a n o p a r t i c l e s , o r

a n h y d r o u s

c o m p o s i t i o n s . A

p r e f e r r e d t o p i c a l

c a r r i e r

i s a cream o r a

poWder.

[ 0 0 7 5 ]

The

p r e s e n t i n v e n t i o n

a l s o d i s c l o s e s a method o f

t r e a t i n g a

s k i n c o n d i t i o n

c o m p r i s i n g

t o p i c a l l y

a p p l y i n g

t o t h e

s k i n , f o r a

p e r i o d o f t i m e

and n an amount s u ? i c i e n t t o

e f f e c t

c h a n g e s i n t h e

d e r m i s ,

t h e c o m p o s i t i o n ( s ) l i s t e d a b o v e . The

c o m p o s i t i o n ( s )

may

e

a p p l i e d

t o

an

n d i v i d u a l ’ s

s k i n W i t h i n

a

r e g i o n i n

Which

r e v e n t i o n a n d / o r t r e a t m e n t i s

d e s i r e d .

The

c o m p o s i t i o n ( s )

may

be

p p l i e d

on

n

a s - n e e d e d b a s i s o r p e r i

o d i c a l l y o v e r an extended p e r i o d of

t i m e , s u c h

a s , e .

g . ,

at

every d i a p e r change,

at l e a s t once

a

day

o r a t

l e a s t once

a

w e e k . t

i s p r e f e r r e d o a p p l y t h e

c o m p o s i t i o n ( s )

o f t h e p r e s e n t

i n v e n t i o n as f r e q u e n t l y a s p o s s i b l e .

[ 0 0 7 6 ]

A c c o r d i n g

t o

a n e m b o d i m e n t , t h e

pH

f t h e compo

s i t i o n i s a b o u t 4 . 5 t o a b o u t 8 . According to a p r e f e r r e d

e m b o d i m e n t , t h e

pH

f t h e c o m p o s i t i o n

i s a b o u t 5 t o a b o u t

6 .

A c c o r d i n g t o a

more

p r e f e r r e d e m b o d i m e n t , t h e pH o f t h e

c o m p o s i t i o n i s

a b o u t

5 t o

a b o u t

5 . 5 .

According

to

a most

p r e f e r r e d e m b o d i m e n t , t h e pH o f t h e c o m p o s i t i o n i s a b o u t

5 . 5 .

8/16/2019 Us 20130004580

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US 2013/0004580

A 1

EXAMPLES

[ 0 0 7 7 ]

The f o l l o w i n g

e x a m p l e s a r e p r e s e n t e d t o i l l u s t r a t e

t h e i n v e n t i o n . A s

i l l u s t r a t i o n s

t h e y a r e n o t i n t e n d e d t o l i m i t

t h e s c o p e

o f

t h e i n v e n t i o n . A l l q u a n t i t i e s

a r e i n w e i g h t p e r

c e n t .

E x a m p l e 1

The E f f e c t

o f

C a t i o n s

on r y p s i n

I n h i b i t i o n

[ 0 0 7 8 ]

The o b j e c t i v e

o f

t h e f o l l o w i n g

e x p e r i m e n t

was

o

d e t e r m i n e w h e t h e r

and

to

what

e x t e n t t h e

f o l l o w i n g c a t i o n s

i n h i b i t e d t r y p s i n a c t i v i t y i n a n i n v i t r o a s s a y .

[ 0 0 7 9 ]

The n h i b i t i o n o f t r y p s i n - i n d u c e d

c l e a v a g e

o f

a ?uo

r e s c e n t c a s e i n p e p t i d e was measured u s i n g

t h e

EnZChekTM

p r o t e a s e a s s a y k i t ,

f o l l o w i n g

m a n u f a c t u r e r ’ s

i n s t r u c t i o n s

( E n Z C h e k T M P r o t e a s e A s s a y

K i t s

P r o d u c t I n f o r m a t i o n ,

R e v i s e d ;

M o l e c u l a r

P r o b e s ,

E u g e n e O r e g . ) .

B r i e ? y ,

t e s t

m a t e r i a l s

a t d i f f e r e n t c o n c e n t r a t i o n s were p r e p a r e d i n 1x

p h o s p h a t e

b u f f e r e d

s a l i n e

( P B S ,

pH

. 4 ) .

T r y p s i n

w o r k i n g

s o l u t i o n

( S i g m a ,

S t . L o u i s , M o . ,

u n i t / m L )

w a s p r e p a r e d i n

d i g e s t i o n b u f f e r p r o v i d e d i n t h e a s s a y

k i t .

S t o c k s o l u t i o n o f

BODIPY FL

a s e i n

( t r y p s i n s u b s t r a t e , mg/mL) was r e p a r e d

b y

a d d i n g

0 . 2 mL

o

t h e s u b s t r a t e v i a l s ( p r o v i d e d i n

k i t ) ,

a n d

t h e

? n a l

s u b s t r a t e w o r k i n g s o l u t i o n ( 1 0

m i c r o g r a m / m L )

was

p r e p a r e d b y d i l u t i o n i n d i g e s t i o n b u f f e r ( p H

7 . 8 ) .

F o l l o w i n g

i n c u b a t i o n o f t r y p s i n , w i t h o r

w i t h o u t

t h e t e s t m a t e r i a l s , w i t h

t h e

BODIPY u o r e s c e n t c a s e i n s u b s t r a t e , a t room

tempera

t u r e f o r one

o u r ,

? u o r e s c e n c e

was measured

e x c i t a t i o n 485

n m / e m i s s i o n 5 3 0

nm)

u s i n g a

S p e c t r a M a x m i c r o t i t e r

p l a t e

r e a d e r

( M o l e c u l a r D e v i c e s C o r p o r a t i o n , S u n n y v a l e , C a l i f . )

a n d

S o f t m a x

P r o

s o f t w a r e

( M o l e c u l a r D e v i c e s C o r p o r a t i o n ) .

E a c h e x p e r i m e n t was p e r f o r m e d i n t h r e e r e p l i c a t e s . The

r e s u l t s o f

t h e e x p e r i m e n t a r e

shown n T a b l e

1 . I n g e n e r a l , t h e

r e s u l t s

demonstrate

h a t

d i v a l e n t c a t i o n s a r e

more

o t e n t

t h a n

m o n o v a l e n t c ati o n s

i n t h e i r

t r y p

i n

i n h i b i t o r y a c t i v i t y .

TABLE

M o l e c -

C o n c e n - C o n c e n - %

r y p s i n

C h e m i c a l M o l e c u l a r

u l a r

t r a t i o n % t r a t i o n

i n h i b i t o r y

name

f o r m u l a

w e i g h t ( g / m L )

(mM)

a c t i v i t y

( I I )

c h l o r i d e C u C l 2 ° ( H 2 O ) 2 1 7 0 . 4 8

0 . 0 1

% 0 . 5 9 6 4 . 9 8

d i h y d r a t e 0.10 5 . 8 7 9 9 . 3 4

1%

5 8 . 6 6

1 0

8 . 3 4

J a n .

3 ,

2 0 1 3

TABLE

- c o n t i n u e d

M o l e c - C o n c e n - C o n c e n - % r y p s i n

C h e m i c a l M o l e c u l a r u l a r t r a t i o n % t r a t i o n i n h i b i t o r y

name

f o r m u l a w e i g h t (gmL)

(mM)

a c t i v i t y

Z i n c

c h l o r i d e ZnCl2

1 3 6 . 3

0.01

0 . 7 3

4 2 . 9 9

0 .1 0

7 . 3 4 8 8 . 5 1

1%

7 3 . 3 7 105.87

I r o n

( I I ) F e C l 2 ° ( H 2 O ) 4 1 9 8 . 8 1 0.01 0 . 5 0 — 3 . 2 4

c h l o r i d e

0 .1 0

5 . 0 3 6 4 . 8 4

t e t r a h y d r a t e 1

5 0 . 3

1 1 2 . 2 3

Calcium

CaCl2

1 1 0 . 9 8 0.01 0 . 9 — 1 0 . 6 3

c h l o r i d e

0 .1 0

9 . 0 1 4 6 . 5 2

1%

9 0 . 1 1 7 1 . 3 5

S t r o n t i u m S r C l 2 1 5 8 . 5 3 0.01 0 . 6 3 6 . 5 1

c h l o r i d e

0 .1 0

6 . 3 1

1 1 . 7 8

1%

6 3 . 0 8

4 7 . 0 5

Magnesium MgCl2

9 5 . 2 1 0.01 1 . 0 5 3 . 2 9

c h l o r i d e

0 .1 0 1 0 . 5 1 2 . 7 2

1% 1 0 5 . 0 3 3 9 . 3 2

Sodium NaCl 5 8 . 4 4 0. 01

1 . 7 1

—7.57

c h l o r i d e

0 .1 0

1 7 . 1 1 — 4 . 1 5

1%

171.12 1 0 . 8 8

Lithium

L i C l 4 2 . 3 9

0. 01

2 . 3 6

8 . 7 1

c h l o r i d e

0 .1 0

2 3 . 5 9

7 . 5 6

1% 2 3 5 . 9

—0.56

S i l v e r

AgNO3 1 6 9 . 8 7

0. 01

0 . 5 9 — 2 . 4 3

n i t r a t e

0 .1 0

5 . 8 9

2 . 9 1

1% 5 8 . 8 7

5 . 3 7

The r e s u l t s

d e m o n s t r a t e

t h a t , o v e r a l l , d i v a l e n t c a t i o n s

a r e

more p o t e n t t h a n

m o n o v a l e n t c a t i o n s i n

t r y p s i n

i n h i b i t o r y

a c t i v i t y . I n p a r t i c u l a r , t h e r e s u l t s o f E x a m p l e 1 d e m o n s t r a t e

t h a t w i t h r e g a r d t o t r y p s i n

i n h i b i t i o n , a c t i v i t y

i s i n t h e f o l l o w

i n g

o r d e r :

Z n + 2 > F e + 2 > C a + 2 > S r + 2 > M g + 2 .

T h e

r e s u l t s

a l s o

show t h a t , o v e r a l l , a l l o f t h e d i v a l e n t c a t i o n s

h a v e h i g h e r

a c t i v i t y

t h a n t h e m o n o v a l e n t c a t i o n s ,

N a “ ,

Ag“, a n d L i “ .

E x a m p l e 2

The

E f f e c t

o f

S a l t

Form

o f

t h e

Same

i v a l e n t C a t i o n

on r y p s i n I n h i b i t i o n

[ 0 0 8 0 ]

The o b j e c t i v e

o f

t h e f o l l o w i n g

e x p e r i m e n t was t o

d e t e r m i n e

whether and

o

what e x t e n t d i f f e r e n t s a l t forms of

t h e same

d i v a l e n t

c a t i o n had on n h i b i t i o n o f t r y p s i n a c t i v i t y

i n

an

i n v i t r o a s s a y .

[ 0 0 8 1 ] T r y p s i n

i n h i b i t o r y

a c t i v i t y w a s a n a l y Z e d

a s

d e s c r i b e d i n Example a n d

t h e

e f f e c t o f

d i f f e r e n t

s a l t f o r m s

of t h e

same

c a t i o n s

on

t h e t r y p s i n i n h i b i to r y

a c t i v i t i e s

was

a s s e s s e d .

T a b l e 2

s h o w s t h e

m o l e c u l a r w e i g h t s , c o n c e n t r a t i o n

a n d

t r y p s i n i n h i b i t o r y a c t i v i t y

o f

t e s t e d

c o m p o u n d s .

TABLE

%

r y p s i n

M o l e c u l a r

% e i g h t i n h i b i t o r y

C h e m i c a l

name

M o l e c u l a r f o r m u l a w e i g h t

(gmL)

mM

a c t i v i t y

Z i n c c h l o r i d e ZnCl2 1 3 6 . 3 2 0.01 0 . 7 3 4 1 . 2 5

0 .1 0

7 . 3 4 8 7 . 6 9

1%

73 3 6 9 8

. 7 5

Zinc a c e t a t e

Zn(C2H3O2)2°(H2O)2

2 1 9 . 5 0. 01

0 . 4 6 3 6 . 2 2

d i h y d r a t e 0.10 4 . 5 6 6 8 . 9 2

Z i n c g l u c o n a t e

Z i n c

l a c t a t e

1% 45.56

9 3 . 8

2 1 1 0 6 1 1 1 1 0 7 ) 2 4 5 5 . 6 8 0.01 0 . 2 2 2 5 . 3 3

0 .1 0

2 . 1 9 4 0 . 9 3

1%

21.94 8 5 . 5 2

2 1 1 0 6 1 1 1 1 0 7 ) 2 2 4 3 . 5 5 0.01 0 . 4 1

2 3 . 0 1

0 .1 0 4 . 1 1

3 6 . 3 6

1%

41.06 8 7 . 9 5

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US 2 0 1 3 / 0 0 0 4 5 8 0

A1

J a n .

3 ,

2 0 1 3

TABLE - c o n t i n u e d

%

r y p s i n

M o l e c u l a r

% w e i g h t

i n h i b i t o r y

C h e m i c a l name

M o l e c u l a r f o r m u l a w e i g h t

(gmL)

mM

a c t i v i t y

C o p p e r

( 1 1 )

C u ( C 2 H 3 O 2 ) 2 ° ( H 2 O ) 1 9 9 . 6 5

0 . 0 1 % 0 . 5 0 6 6 . 5 3

a c e t a t e

0 .1 0

5 . 0 1

95.17

m o n o h y d r a t e

C o p p e r

( 1 1 ) C u ( C 1 2 H 2 2 O 1 4 ) 4 5 3 . 8 0.01 6 . 3 2 1 4 . 3 8

g l u c o n a t e 0.10 6 3 . 2 2 6 5 . 5 2

Calcium

c h l o r i d e CaCi2 1 1 0 . 9 8 0.10 9 . 0 1 3 3 . 4 1

1%

9 0 . 1 1 7 1 . 3 9

Calcium

a c e t a t e Ca(C2H3O2)2°(H2O)

1 7 6 . 1 7

0. 10 5 . 6 8 1 4 . 3 8

monohydrate 1 5 6 . 7 6 6 5 . 5 2

Magnesium

MgCl2

9 5 . 2 1

0.10

1 0 . 5 3 9 . 0 2

c h l o r i d e

Magnesium Mg(C2H3O2)2°(H2O)4

2 1 4 . 4 5

0.10

4 . 6 6 - 3 . 0 9

a c e t a t e

t e t r a h y d r a t e

The

r e s u l t s

d e m o n s t r a t e

t h a t

d i f f e r e n t s a l t

f o r m s

o f s i m i l a r TABLE

c a t i o n s

h a v e

d i f f e r e n t

p o t e n c i e s

i n

t e r m s

o f

t r y p s i n

i n h i b i t o r y

a c t i v i t y ‘ H y p o t h e t i c a l l y

y n e r g i s t i c

c a l c u l a t e d a d d i -

a c t i v i t y

[ 0 0 8 2 ]

A s

s h o w n i n T a b l e 2 , Z n 2 + i n t h e c h l o r i d e s a l t f o r m f a “ ‘ £ 1 1 m

“ 1 :

V a i u g l ’ f

{ 1 6 1 M 1 5 1

s o

. . . . .. . . 3.0116 3C

S3. 3 . 1 1 C ypO 61

h a s t h e h l g h e s t t r y p s 1 n 1 n h 1 b 1 t 1 o n a c t 1 v 1 t y , f o l l o w e d

by

t h e S a m p l e s t r y p s i n i n h i b i t i o n a c t i v i t i e s

( )

a d d i t i v e V a l u e

a c e t a t e , g l u c o n a t e

a n d

l a c t a t e s a l t f o r m s when a h b r a t e d b y

molar c o n c e n t r a t i o n s .

T a b l e 2

a l s o shows t h a t , f o r a

g i v e n T a l c

8 ' 5 5

8 ' 5 5 0 ' 0 0

. . . . . . . . . . .

CaCl2 17.95

26.43 26.50 -0.26

d 1 v a l e n t ~ c a t 1 o n , ~ a s 1 m 1 l a r o r d e r i n g o f a n 1 o n s 1 n t r y p s 1 n 1 n h 1 b 1 - M g c l 2

4 7 5

2 0 0 5 1 3 3 0 5 0 7 5

t o r y a c t 1 v 1 t 1 e s 1 s o b s e r v e d . T h e r e f o r e ,

chooslng

t h e amon

F e C l 2 ° ( H 2 O ) 4

6 5 . 0 7 6 4 . 5 3

7 3 . 6 2

- 1 2 . 3 5

c o u l d

a f f e c t t h e t r ' y p s i n i n h i b i t o r y

a c t i v i t y

o f t h e d i v a l e n t C a ( C 2 H 3 O 2 ) 2 ° ( H 2 O ) 1 5 - 3 1 2 4 - 6 2 3 - 8 6

3 ' 1 0

. . . . .

Zn(C2H3O2)2°(H2O)2 62.84

60.18

71.39 -15.70

c a t 1 o n - c o n t a 1 n 1 n g

compos1t1on.

Z H C I Z

81m 7796

8962

_ 1 3 _ 0 1

Mg(C2H3O2)2°(H2O)4 0 . 8 3 1 7 . 8 3 9 . 3 8 9 0 . 0 9

E x a m p l e 3

[ 0 0 8 3 ]

The o b j e c t i v e

o f

t h e f o l l o w i n g

e x p e r i m e n t

was

o

d e t e r m i n e

whether

and

o

what x t e n t a d d i n g a d i v a l e n t c a t i o n

to t a l c i n h i b i t e d

t r y p s i n a c t i v i t y

i n an

i n v i t r o a s s a y . The

i n h i b i t i o n o f

t r y p s i n - i n du c e d c l e a v a g e o f

a ? u o r e s c e n t

c a s e i n

p e p t i d e was

m e a s u r e d u s i n g t h e EnZChekTM

r o t e a s e

a s s a y

k i t , f o l l o w i n g m a n u f a c t u r e r ’ s i n s t r u c t i o n s ( E n Z C h e k T M P r o

t e a s e A s s a y

K i t s P r o d u c t I n f o r m a t i on , R e v i s e d ; M o l e c u l a r

P r o b e s , E u g e n e

O r e g . ) .

B r i e ? y ,

m i x t u r e s

o f

t a l c

s t o c k s o l u

t i o n s

w i t h

( 1 ) d i v a l e n t c a t i o n s a l t s a t d i f f e r e n t c o n c e n t r a t i o n s

( a s i n d i c a t e d i n T a b l e 3 )

a n d

( 2 ) magnesium c h l o r i d e

a t d i f

f e r e n t

c o n c e n t r a t i o n s

( a s

i n d i c a t e d

i n T a b l e 4 )

w e r e

p r e p a r e d

i n

1 x p h o s p h a t e b u f f e r e d s a l i n e ( P B S , pH . 4 ) . T r y p s i n

w o r k

i n g s o l u t i o n ( S i g m a , S t . L o u i s , M o . , u n i t / m L ) w a s p r e p a r e d i n

d i g e s t i o n b u f f e r p r o v i d e d i n t h e a s s a y k i t . S t o c k s o l u t i o n o f

BODIPY

FL

a s e i n

( t r y p s i n

s u b s t r a t e ,

mg/mL)

was

r e p a r e d

b y

a d d i n g

0 . 2

mL

o

t h e s u b s t r a t e v i a l s ( p r o v i d e d i n k i t ) ,

a n d

t h e

? n a l

s u b s t r a t e w o r k i n g s o l u t i o n ( 1 0 m i c r o g r a m / m L ) was

p r e p a r e d b y d i l u t i o n i n d i g e s t i o n b u f f e r ( p H

7 . 8 ) . F o l l o w i n g

i n c u b a t i o n o f t h e t r y p s i n , w i t h

o r

w i t h o u t t h e t e s t

m a t e r i a l s ,

with t h e

BODIPY

u o r e s c e n t

c a s e i n s u b s t r a t e , a t

room

tem

p e r a t u r e f o r

one o u r ,

? u o r e s c e n c e was measured

e x c i t a t i o n

4 8 5 n m / e m i s s i o n 5 3 0 nm) u s i n g a S p e c t r a M a x m i c r o t i t e r

p l a t e r e a d e r ( M o l e c u l a r D e v i c e s C o r p o r a t i o n , S u n n y v a l e ,

C a l i f . ) a n d S o f t m a x P r o s o f t w a r e ( M o l e c u l a r D e v i c e s C o r p o

r a t i o n ) .

E a c h e x p e r i m e n t

w a s p e r f o r m e d

i n

t h r e e

r e p l i c a t e s .

The e s u l t s o f t h e e x p e r i m e n t s

a r e

shown n T a b l e 3 and a b l e

4 .

A l l t e s t e d

m a t e r i a l s were t 0. 1 g / m L ) .

* H y p o t h e t i c a l a d d i t i v e v a l u e was

a l c u l a t e d b y

a d d i n g % r y p s i n i n h i b i t i o n o f t h e s a l t a l o n e

a n d

t a l c

a l o n e

( 8 . 5 5 % ) .

**

y n e r g i s t i c

a c t i v i t y was a l c u l a t e d b y s u b t r a c t i n g t h e h y p o t h e t i c a l

ad d iti ve v a l u e

f r o m

t h e

e x p e r i m e n t a l l y m e a s u r e d “ w i t h

t a l c

v a l u e ” t h e n

d i v i d i n g

by

he h y p o t h e t i c a l ad d iti ve

v a l u e .

[ 0 0 8 4 ]

The

s e o f a l c

i s s o o t h i n g

t o d i a p e r r a s h .

The

e s u l t s

s u g g e s t

t h a t a f o r m u l a t i o n

c o n t a i n i n g

magnesium

d i v a l e n t

c a t i o n a n d t a l c c o u l d n o t o n l y

p r o v i d e

s o o t h i n g

r e l i e f b u t

a l s o

i n h i b i t t h e p r o g r e s s i o n

o f h e r a s h b y i n h i b i t i n g t r y p s i n a c t i v

i t y .

TABLE

H y p o t h e t i c a l l y

% y n e r g i s t i c

c a l c u l a t e d a d d i -

a c t i v i t y

S a l t With

t i v e v a l u e of

r e l a t i v e t o

a l o n e 1

a l c

s a l t a n d

T a l c

h y p o t h e t i c a l

S a m p l e s t r y p s i n i n h i b i t i o n a c t i v i t i e s

( % )

a d d i t i v e

v a l u e

no Mg Cl2 0 3 . 4 2 3 . 4 2 0 . 0 0

0. 1 MgCl2 9 . 0 2

2 5 . 4 3 1 2 . 4 4 1 0 4 . 4 2

0.5 MgCl2 1 9 . 5 1 3 2 . 8 4 2 2 . 9 3 4 3 . 2 2

1 MgCl2

2 2 . 1 3

4 0 . 4 2

2 5 . 5 5 5 8 . 2 0

A l l

c o n c e n t r a t i o n s l i s t e d a r e

g/mL.

[ 0 0 8 5 ]

The r e s u l t s

d e m o n s t r a t e

t h a t t a l c a n d magnesium

c h l o r i d e a l o n e h a v e m o d e r a t e

to

low

l e v e l s

o f

t r y p s i n

i n h i b i

t o r y a c t i v i t y ,

a n d t h a t t h e c o m b i n a t i o n o f a l c

a n d

magnesium

c h l o r i d e had a

s y n e r g i s t i c

i n c r e a s e i n t r y p s i n

i n h i b i t i o n .

E x a m p l e 4

T r y p s i n

I n h i b i t i o n

w i t h Z i n c O x i d e C o n t a i n i n g

F o r m u l a t i o n

[ 0 0 8 6 ] The o b j e c t i v e

o f

t h e f o l l o w i n g e x p e r i m e n t was t o

determine whether and t o what e x t e n t adding an a c i d t o a

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A 1

f o r m u l a t i o n

c o n t a i n i n g Z i n c

o x i d e

had on n h i b i t i o n

o f

t r y p

s i n

a c t i v i t y

i n a n i n

v i t r o

a s s a y .

The

n h i b i t i o n

o f t r y p s i n - i n d u c e d

c l e a v a g e o f

a ? u o r e s c e n t c a s e i n p e p t i d e Was measured

s i n g

t h e EnZChekTM

r o t e a s e a s s a y k i t ,

f o l l o w i n g m a n u f a c t u r e r ’ s

i n s t r u c t i o n s

( E n Z C h e k T M

P r o t e a s e

A s s a y

K i t s

P r o d u c t

I n f o r

m a t i o n , R e v i s e d ; M o l e c u l a r

P r o b e s , E u g e n e

O r e g . ) .

B r i e ? y ,

m i x t u r e s o f Z i n c o x i d e

and

a c i d

a t

d i f f e r e n t c o n c e n t r a t i o n s

W e r e p r e p a r e d in

l > < P B S

(pH . 4 )

a n d

t h e i r pH Was e c o r d e d

a s

l i s t e d i n

t h e T a b l e s 5 - 8 . T r y p s i n W o r k i n g s o l u t i o n

( S i g m a ,

S t . L o u i s , M o . , u n i t /m L ) W a s p r e p a r e d i n d i g e s t io n b u f f e r

p r o v i d e d i n

t h e

a s s a y k i t . S t o c k s o l u t i o n o f BODIPY FL

c a s e i n ( t r y p s i n

s u b s t r a t e ,

mg/mL)

a s

p r e p a r e d

b y a d d i n g 0 . 2

mL o t h e

s u b s t r a t e

v i a l s ( p r o v i d e d

i n

k i t ) , a n d t h e ? n a l

s u b s t r a t e

W o r k i n g s o l u t i o n ( 1 0 m i c r o g r a m / m L ) W a s p r e p a r e d

b y d i l u t i o n i n d i g e s t i o n b u f f e r ( p H 7 . 8 ) . F o l l o w i n g i n c u b a t i o n

o f t h e t r y p s i n ,

W i t h

o r W i t h o u t t h e t e s t m a t e r i a l s

( 5 0 p l ) , W i t h

t h e BODIPY u o r e s c e n t c a s e i n s u b s t r a t e , a t room

tempera

t u r e

f o r

one

o u r , ?uorescence

Was

measured e x c i t a t i o n

485

n m / e m i s s i o n 5 3 0

nm)

u s i n g a

S p e c t r a M a x m i c r o t i t e r

p l a t e

r e a d e r

( M o l e c u l a r D e v i c e s

C o r p o r a t i o n ,

S u n n y v a l e ,

C a l i f . )

a n d S o f t m a x P r o s o f t W a r e ( M o l e c u l a r

D e v i c e s

C o r p o r a t i o n ) .

E a c h

e x p e r i m e n t

Was e r f o r m e d i n t h r e e r e p l i c a t e s .

[ 0 0 8 7 ] F o r p r e p a r a t i o n s , s a m p l e s Were i l u t e d i n l > < P B S a t

1

: 1 0

r a t i o

and

u b j e c t e d t o

c e n t r i f u g a t i o n a t

1 3 , 2 0 0 rpm o r 1

m i n u s i n g E p p e n d o r f C e n t r i f u g e ( M o d e l 5 4 15 D , E p p e n d o r f

AG,

Hamburg, Germany)

t o

remove t h e s o l i d

f o r m u l a t i o n

i n g r e d i e n t s . T h e

s u p e r n a t a n t s

( 5 0

p l )

W e r e u s e d f o r t r y p s i n

i n h i b i t o r y

a c t i v i t y

a n a l y s e s as d e s c r i b e d a b o v e . F o r p r e p a r a

t i o n s , s a m p l e s Were d i l u t e d i n 1 0 , o r

20 o r

40%

t h a n o l

and

s u b j e c t e d t o

c e n t r i f u g a t i o n a t

1 3 , 2 0 0

rpm

f o r 1 min u s i n g

E p p e n d o r f e n t r i f u g e

( M o d e l

5 4 1 5 D ,

E p p e n d o r f

A G , Ham

b u r g , Germany) o remove h e

s o l i d

f o r m u l a t i o n

i n g r e d i e n t s .

The

s u p e r n a t a n t s ( 5 0

p l )

W e r e u s e d f o r t r y p s i n i n h i b i t i o n

a s s a y . P o s s i b l e

e f f e c t s

o f e t h a n o l s o l u t i o n ( 1 0 , 20 o r 40%

e t h a n o l ) on

r y p s i n

i n h i b i t i o n a c t i v i t y Were

e s t e d

i n t h e

same

s t u d y

a n d f o u n d n o

s i g n i ? c a n t i m p a c t .

The

r e s u l t s

o f

t h e

e x p e r i m e n t s

a r e

shoWn

n

T a b l e s 5 , 6 , 7 a n d 8 .

P r e p a r a t i o n s 1 a n d

2 .

Z i n c O x i d e C o n t a i ni n g F o r m u l a t i o n

W i t h o u t

o r

W i t h V a r y i n g

C o n c e n t r a t i o n s o f A c e t i c A c i d .

[ 0 0 8 8 ]

TWo

r e p a r a t i o n s

of

h e

same

o r m u l a t i o n

Were r e

p a r e d on i f f e r e n t

d a y s .

The a s e

f o r m u l a t i o n

f o r e a c h p r e p a

r a t i o n

c o n t a i n e d t h e i n g r e d i e n t s l i s t e d beloW and

Was p r e

p a r e d a s f o l l o W s : 1 . P r e p a r e

1

N c e t i c a c i d

s o l u t i o n . 2 .

I n a

b e a k e r

Weigh h e amount o f

W a t e r

needed

o r

e a c h

f o r m u l a

t i o n a n d b e g i n h e a t i n g t o 6 5 - 7 0 0 C . 3 . Add

e t h y l

a n d

p r o p y l

p a r a b e n a t

7 0 0

C . and

e t

i t s t i r

f o r

5

m i n u t e s

and

e d u c e

t h e

h e a t . 4 .

Add

h e a p p r o p r i a t e amount o f a c e t i c a c i d

( c o n t r o l

c o n t a i n s

no

a c e t i c a c i d ) . 5 . A d d

i n c

o x i d e

and

e t i t r e a c t f o r

5

m i n u t e s a t 5 5 - 6 0 0

C .

6 . Add

i n e r a l

o i l

and

e t r o l a t u m

to

t h e m i x t u r e

and a f t e r

t h e

p e t r o l a t u m m e l t s

t u r n t h e h e a t o f f

and

e t

t h e

m i x t u r e c o o l W i t h c o n s t a n t

s t i r r i n g . 7 .

Measure

h e

p H .

[ 0 0 8 9 ] T h e

t r y p s i n i n h i b i t o r y

a c t i v i t y

f o r t h e

t W o p r e p a r a

t i o n s

a r e shoWn n

T a b l e 6 .

The

r e p a r a t i o n s e x h i b i t e d

i n s t a

b i l i t y

o v e r t i m e .

P r e p a r a t i o n s 1 a n d 2

1 0

i n c o x i d e

a c e t i c

a c i d a t v a r y i n g c o n c e n t r a t i o ns ( n o

a c i d ; 0 . 1 — 0 . 6 % )

0 . 1 8 %

m e t h y l p a r a b e n

0 . 0 2 % p r o p y l

p a r a b e n

19.39 i n e r a l o i l

J a n .

3 ,

2 0 1 3

- c o n t i n u e d

P r e p a r a t i o n s

1 a n d

2

2 5 . 8 6 %

p e t r o l a t u m

W a t e r

a t

v a r y i n g c o n c e n t r a t i o n s ( 3 2 . 3 4 — 3 4 % ) .

P r e p a r a t i o n s 3 ,

4

a n d

5 . T r y p s i n

I n h i b i t o r y

A c t i v i t y o f

F o r m u l a t i o n

C o n t a i n i n g

( 1 )

Z i n c

O x i d e

a n d

No i t r i c A c i d ;

( 2 ) Z i n c

O x i d e a n d 0.1 C i t r i c

A c i d ; ( 3 ) Z i n c

O x i d e a n d

0. 5

C i t r i c A c i d .

[ 0 0 9 0 ] P r e p a r a t i o n s

3 ,

4 , a n d

5 ,

t h e r e s u l t s f o r W h i c h a r e

p r e s e n t e d

i n T a b l e s

7

a n d 8

b e l o W , c o n t a i n ed

t h e i n g r e d i e n t s

l i s t e d

beloW and Were

r e p a r e d

a s f o l l o W s :

P r e p a r a t i o n

3

[ 0 0 9 1 ]

I n g r e d i e n t % t / W t f o r 200 g

O i l C12-15

a l k y l b e n z o a t e 5 . 5 3 1 1 . 0 5

p h a s e I s o p r o p y l P a l m i t at e 4 . 4 2

8 . 8 4

Zinc S t e a r a t e

0 . 5 0 1 . 0 0

D i c a p r y l y l c a r b o n a t e

5

. 5 3 1 1 . 0 5

P o l y g l y c e r y l - 2 3 . 3 2 6 . 6 4

d i p o l y h y d r o x y s t e a r a t e

P o l y g l y c e r y l - 3 d i i s o s t e a r a t e 2 . 2 1 4 . 4 2

L a n o l i n 4 . 0 0 8 . 0 0

( 9 9 . 9 8 ) / b u t y l h y d r o x y t o l u e n e

( 0 . 0 2 )

( “ l a n o l i n

B H T ” )

Zinc

Oxide 1 3 . 0 0 2 6 . 0 0

Water W a t e r ,

D e r n i n e r a l i z e d 5 9 . 4 3

1 1 8 . 8 6

p h a s e Disodium EDTA 0 . 2 0 0 . 4 0

Magnesium S u l f a t e

1 . 0 0

2 . 0 0

H e p t a h y d r a t e

P h e n o x y e t h a n o l

0 . 5 0 1 . 0 0

M e t h y l p a r a b e n 0 . 3 0 0 . 6 0

P r o p y l p a r a b e n

0 . 0 7 0 . 1 4

100.00

200.00

[ 0 0 9 2 ] O i l P h a s e

[ 0 0 9 3 ] 1 . Combine 1 2 - 1 5 a l k y l b e n Z o a t e a n d I s o p r o p y l

P a l m i t a t e

i n a n

a p p r o p r i a t e l y s i Z e d

b e a k e r

and r i n g t o

1 2 0 - 1 3 0 0

C .

Mix t 1 0 0 - 3 0 0 rpm n t i l homogeneous.

[ 0 0 9 4 ]

2 .

A d d i n c S t e a r a t e

a t

1 2 0 - 1 3 0 0 C .

M i x

t

100

3 0 0 rpm

n t i l

h o m o g e n e o u s .

[ 0 0 9 5 ] 3 . AlloW i x t u r e to c o o l t o 9 5 - 1 0 5 o C . A d d Dica

p r y l y l

c a r b o n a t e a n d mix t 1 0 0 - 3 0 0 rpm n t i l homoge

neous.

[ 0 0 9 6 ]

4 .

Add

o l y g l y c e r y l - 2 d i p o l y h y d r o x y s t ea r a t e

t o

t h e

m i x t u r e

a t 90-105o

C .

M i x a t 100-300 rpm

u n t i l

h o m o g e n e o u s .

[ 0 0 9 7 ] 5 .

AlloW

m i x t u r e t o c o o l

to 8 5 - 9 5 0

C . and add

P o l y g l y c e r y l - 3 d i i s o s t e a r a t e .

Mix t

1 0 0 - 3 0 0 rpm n t i l

h o m o g e n e o u s .

[ 0 0 9 8 ] 6 . A d d a n o l i n BHT

t

8 5 - 9 5 0 C .

M i x t

200-500

rpm n t i l

h o m o g e n e o u s .

[ 0 0 9 9 ]

7 .

A d d i n c Oxide t 8 5 - 9 5 0 C . Mi x t 200-500

rpm n t i l

h o m o g e n e o u s .

[ 0 1 0 0 ] 8 . HomogeniZe

m i x t u r e a t

5 0 0 0 - 6 0 0 0 rpm

o r 5

m i n u t e s .

[ 0 1 0 1 ] 9 .

Remove

i x t u r e f r o m

h o m o g e n i Z e r

a n d k e e p

at 85-900 C .

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A 1

[ 0 1 0 2 ] W a t e r

P h a s e

[ 0 1 0 3 ] 1 0 . I n a s e p a r a t e b e a k e r , combine ater

and i s o

dium EDTA a t

40-500

C . M i x a t 100-300 rpm u n t i l

h o m o g e n e o u s .

[ 0 1 0 4 ]

1 1 .

B r i n g m i x tu r e

t o

5 0 - 6 0 0

C .

a n d a d d

Magne

sium S u l f a t e

H e p t a h y d r a t e . M i x t 1 0 0 - 3 0 0 rpm

n t i l

h o m o g e n e o u s .

[ 0 1 0 5 ] 1 2 . Add

h e n o x y e t h a n o l

a t

5 0 - 6 0 0 C . M i x

t

1 0 0

3 0 0

rpm

n t i l

h o m o g e n e o u s .

[ 0 1 0 6 ] 1 3 .

Add e t h y l p a r a b e n a t

5 0 - 6 0 0

C . Mix t 1 0 0

3 0 0 rpm

n t i l

h o m o g e n e o u s .

[ 0 1 0 7 ]

1 4 .

Add r o p y l p a r a b e n a t

5 0 - 6 0 0

C . Mix t 1 0 0

3 0 0

rpm

n t i l h o m o g e n e o u s .

[ 0 1 0 8 ] 1 5 . B r i n g m i x t u r e t o 8 5 - 9 0 0 C .

[ 0 1 0 9 ]

E m u l s i o n

P h a s e

[ 0 1 1 0 ] 1 6 . S l o W l y a d d

W a t e r

P h a s e

c o n t e n t s

i n t o O i l

Phase t 8 5 - 9 0 0 C . I n c r e a s e mix s p e e d a s n e c e s s a r y .

M i x

f o r 20

i n u t e s .

[ 0 1 1 1 ]

1 7 .

HomogeniZe

m i x t u r e

a t

8 5 - 9 0 0

C .

a t

6000

8000 rpm

o r

5

m i n .

[ 0 1 1 2 ] 1 8 .

Remove

i x t u r e f r o m h o m o g e n i Z er a n d mix

W i t h p r o p e l l e r b l a d e s a t mix s p e e d u s e d i n S t e p 1 6 ( + / —

20 ).

Remove

e a t

and

l l o W t o c o o l

a t

room

tempera

t u r e .

[ 0 1 1 3 ] 1 9 . When

m i x t u r e

c o o l s t o

3 0 - 4 0 0

C . ,

homog

e n i Z e at 6000-8000 rpm

o r

2 m i n .

P r e p a r a t i o n

4

[ 0 1

1 4 ]

I n g r e d i e n t % t / W t f o r 200 g

O i l C12-15

a l k y l b e n z o a t e 5 . 5 3 1 1 . 0 5

p h a s e I s o p r o p y l P a l m i ta t e 4 . 4 2

8 . 8 4

Zinc S t e a r a t e

0 . 5 0 1 . 0 0

D i c a p r y l y l c a r b o n a t e

5

. 5 3 1 1 . 0 5

P o l y g l y c e r y l - 2 3 . 3 2 6 . 6 4

d i p o l y h y d r o x y s t e a r a t e

P o l y g l y c e r y l - 3 d i i s o s t e a r a t e 2 . 2 1 4 . 4 2

L a n o l i n 4 . 0 0 8 . 0 0

( 9 9 . 9 8 ) / b u t y l h y d r o x y t o l u e n e

( 0 . 0 2 )

Zinc

Oxide 1 3 . 0 0 2 6 . 0 0

Water Water

, D e m i n e r a l i z e d 3 9 . 3 3 7 8 . 6 6

p h a s e W a t e r

2 ,

D e m i n e r a l i z e d

2 0 . 0 0 4 0 . 0 0

C i t r i c a c i d

0 . 1 0 0 . 2 0

D i s o d i u r n

EDTA

0 . 2 0 0 . 4 0

Magnesium

S u l f a t e

1 . 0 0

2 . 0 0

H e p t a h y d r a t e

P h e n o x y e t h a n o l 0 . 5 0

1 . 0 0

M e t h y l p a r a b e n

0 . 3 0 0 . 6 0

P r o p y l p a r a b e n

0 . 0 7 0 . 1 4

100.00 200.00

[ 0 1 1 5 ] 0 1 1 P h a s e

[ 0 1 1 6 ] 1 . Combine

1 2 - 1 5 a l k y l b e n Z o a t e a n d I s o p r o p y l

P a l m i t a t e i n a n a p p r o p r i a t e l y

s i Z e d

b e a k e r and r i n g to

1 2 0 - 1 3 0 0

C .

Mix

t

1 0 0 - 3 0 0 rpm n t i l

homogeneous.

[ 0 1 1 7 ]

2 .

A d d i n c S t e a r a t e a t 1 2 0 - 1 3 0 0 C .

M i x

t 100

3 0 0

rpm

n t i l

h o m o g e n e o u s .

[ 0 1 1 8 ] 3 . AlloW i x t u r e t o c o o l t o 9 5 - 1 0 5 o C . Add Dica

p r y l y l c a r b o n a t e a n d mix t 1 0 0 - 3 0 0 rpm

n t i l

homoge

neous.

J a n .

3 ,

2 0 1 3

[ 0 1 1 9 ]

4 .

Add o l y g l y c e r y l - 2 d i p o l y h y d r o x y s t ea r a t e t o

t h e m i x t u r e a t 90-105o C . M i x a t 100-300 rpm

u n t i l

h o m o g e n e o u s .

[ 0 1 2 0 ] 5 . AlloW m i x t u r e t o c o o l

to

8 5 - 9 5 0 C . and add

P o l y g l y c e r y l - 3

d i i s o s t e a r a t e .

Mix

t

1 0 0 - 3 0 0

rpm

n t i l

h o m o g e n e o u s .

[ 0 1 2 1 ] 6 . A d d a n o l i n BHT t 8 5 - 9 5 0 C . M i x t 200-500

rpm n t i l

h o m o g e n e o u s .

[ 0 1 2 2 ] 7 . A d d i n c Oxide t

8 5 - 9 5 0

C . Mi x t 200-500

rpm n t i l

h o m o g e n e o u s .

[ 0 1 2 3 ]

8 . HomogeniZe

m i x t u r e a t

5 0 0 0 - 6 0 0 0 rpm

o r 5

m i n u t e s .

[ 0 1 2 4 ] 9 . Remove i x t u r e

f r o m h o m o g e n i Z e r a n d k e e p

at

85-900 C .

[ 0 1 2 5 ]

W a t e r P h a s e

[ 0 1 2 6 ]

1 0 .

I n

a s e c o n d b e a k e r , combine Water 1 and

C i t r i c

Acid t

4 0 - 5 0 0 C .

M i x t 100-300 rpm

n t i l

homo

g e n e o u s .

[ 0 1 2 7 ]

1 1 .

I n

a t h i r d b e a k e r , combine Water 2 and D i s o

dium

EDTA

t 40-500

C .

M i x a t

100-300

rpm

u n t i l

h o m o g e n e o u s .

[ 0 1 2 8 ] 1 2 .

B r i n g m i x tu r e

t o 5 0 - 6 0 0 C . a n d

a d d Magne

sium S u l f a t e H e p t a h y d r a t e . M i x t 1 0 0 - 3 0 0 rpm

n t i l

h o m o g e n e o u s .

[ 0 1 2 9 ] 13.Add

h e n o x y e t h a n o l

a t

5 0 - 6 0 0

C .

Mix

t 1 0 0

3 0 0 rpm

n t i l

h o m o g e n e o u s .

[ 0 1 3 0 ]

1 4 .

Add

e t h y l p a r a b e n

a t

5 0 - 6 0 0

C . Mix t 1 0 0

3 0 0

rpm

n t i l h o m o g e n e o u s .

[ 0 1 3 1 ] 1 5 .

Add

r o p y l p a r a b e n

a t 5 0 - 6 0 0

C .

M i x

t 1 0 0

3 0 0 rpm n t i l h o m o g e n e o u s .

[ 0 1 3 2 ] 1 6 . Add c o n t e n t s

o f s e c o n d

b e a k e r ( W a t e r

a n d

C i t r i c A c i d

s o l u t i o n ) i n t o t h i r d

b e a k e r a n d

b r i n g m i x t u r e

t o 85-900 C .

[ 0 1 3 3 ]

E m u l s i o n

P h a s e

[ 0 1 3 4 ] 1 7 . S l o W l y

a d d

W a t e r P h a s e c o n t e n t s i n t o O i l

Phase t

8 5 - 9 0 0 C . I n c r e a s e

mix

speed s

n e c e s s a r y . Mi x

f o r 20

m i n u t e s .

[ 0 1 3 5 ] 1 8 . HomogeniZe

m i x t u r e

a t

8 5 - 9 0 0

C .

a t

6000

8000 rpm

o r

5

m i n .

[ 0 1 3 6 ] 1 9 .

Remove i x t u r e f r o m h o m o g e n i Z e r

a n d mix

W i t h p r o p e l l e r b l a d e s a t mix s p e e d u s e d i n S t e p

1 7

( + / —

20 ).

Remove

e a t and

l l o W

t o c o o l a t room tempera

t u r e .

[ 0 1 3 7 ]

2 0 . When m i x t u r e

c o o l s

t o 3 0 - 4 0 0 C . , homog

e n i Z e at 6000-8000 rpm

o r

2 m i n .

P r e p a r a t i o n

5

[ 0 1 3 8 ]

I n g r e d i e n t %

t / W t

f o r 200 g

O i l C12-15 a l k y l b e n z o a t e 5 . 5 3 1 1 . 0 5

p h a s e

I s o p r o p y l

P a l m i t a t e 4 . 4 2 8 . 8 4

Zinc S t e a r a t e 0 . 5 0

1 . 0 0

D i c a p r y l y l c a r b o n a t e

5 . 5 3 1 1 . 0 5

P o l y g l y c e r y l - 2 3 . 3 2 6 . 6 4

d i p o l y h y d r o x y s t e a r a t e

P o l y g l y c e r y l - 3 d i i s o s t e a r a t e

2 . 2 1

4 . 4 2

L a n o l i n 4 . 0 0 8 . 0 0

( 9 9 . 9 8 ) / b u t y l h y d r o x y t o l u e n e

( 0 . 0 2 )

Zinc

Oxide 1 3 . 0 0 2 6 . 0 0

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US 2 0 1 3 / 0 0 0 4 5 8 0

A1

J a n .

3 ,

2 0 1 3

1 0

- C o n t i n u e d

[ 0 1 5 4 ] 1 4 .

Add e t h y l p a r a b e n a t 5 0 - 6 0 0

C .

Mix t 1 0 0

3 0 0 rpm

n t i l

h o m o g e n e o u s .

Ingmdm % WW f o r 2 0 0

g

[ 0 1 5 5 ] 1 5 .

Add

Propylparaben at 50-600 c .

M i x a t 100

W a t e r W a t e r

1 ,

D e m i n e r a l i z e d 3 8 . 9 3 7 7 . 8 6

3 0 0

rpm n t i l

homogeneous'

P 1 1 5 1 5 6

W a t ?

l p m h w r a l i l ? d

2 0 - 0 0 4 0 - 0 0

[ 0 1 5 6 ]

1 6 .

Add

c o n t e n t s

o f s e c o n d

b e a k e r ( W a t e r

a n d

c l m c a c l d

0 ' 5 0 1 ' 0 0

C i t r i c A c i d

s o l u t i o n ) i n t o t h i r d b e a k e r a n d b r i n g m i x t u r e

D i s o d i u m EDTA

0 . 2 0 0 . 4 0

85

9 0 0

C

Magnesium Sulfate

1.00

2 .0 0 to _

H e p t a h y d r a t e

[ 0 1 5 7 ]

E m u l s i o n P h a s e

? w g l o ’ l ‘ y e t h g n o l 8 ' 2 8

( 1 ) 2 8 [ 0 1 5 8 ] 1 7 .

S l o W l y

a d d W a t e r

P h a s e

c o n t e n t s

i n t o

O i l

P r z p y y l g ’ a i l j l ?

0 : 0 7 0 : 1 4

P h a s e

a t

' 8 5 - 9 0 0

C .

I n c r e a s e

m i x

s p e e d s

n e c e s s a r y .

M i x

or

20 minutes.

l o o - 0 0 Z O O - 0 0 [ 0 1 5 9 ]

1 8 .

HomogeniZe m i x t u r e at 8 5 - 9 0 0 C . a t 6000

8000 rpm

o r

5

m i n .

[ 0 1 3 9 ] O i l

p h a s e [ 0 1 6 0 ] 1 9 . Remove i x t u r e f r o m h o m o g e n i Z e r

a n d mix

W i t h p r o p e l l e r b l a d e s a t

mix

s p e e d

u s e d i n

S t e p 1 7 ( + / —

[ 0 1 4 0 ] 1 . Combine 1 2 - 1 5 a l k y l b e n Z o a t e a n d I s o p r o p y l

20%).

Remove

eat and alloW to cool at

room

tempera

P a l m i t a t e i n

an a p p r o p r i a t e l y

s i z e d beaker and r i n g t o m

1 2 0 - 1 3 0 0

C .

Mix

t 1 0 0 - 3 0 0

rpm

n t i l homogeneous. 0 1 6 1 1 1 6 ‘ 2 0 Wh _

1

3 0 4 0 0 C

h

[ 0 1 4 1 ] 2 . Add i I l C

S t e a r a t e

a t 120-1300 c . Mix t 100- l . 1 en mlxmre COO 5 . 1 0 ' Omog'

. en1Ze

a t

6000-8000 rpm or

2 min.

3 0 0

rpm

n t i l

h o m o g e n e o u s .

[ 0 1 4 2 ]

3 .

AlloW

i x t u r e

t o

c o o l

t o

9 5 - 1 0 5 o C .

Add i c a -

R e s u l t s

p r y l y l

c a r b o n a t e a n d mix t 1 0 0 - 3 0 0 rpm n t i l homoge

n e o u s . [ 0 1 6 2 ]

TABLE

T r y p s i n

i n h i b i t o r y

a c t i v i t y o f m i x t u r e s o f

Z i n c o x i d e a n d a c e t i c

a c i d .

No

c e t i c a c i d 0. 01 a c e t i c a c i d 0. 05 a c e t i c a c i d

%

r y p s i n

%

r y p s i n

%

r y p s i n

ZnO i n h i b i t o r y i n h i b i t o r y i n h i b i t o r y

c o n c e n t r a t i o n ( % )

pH a c t i v i t y pH a c t i v i t y pH a c t i v i t y

No Z110 7 . 4 9 0 6 . 8 4

-11.18 + / -

2 . 6 1 4 . 7 5 2 5 . 8 1

+ / -

2 . 1 3

0 .1

Z110 7 . 6 1 — 2 . 6

+ / -

2 . 7 6 7 . 2 8 1 . 8 5 +/-1.34 6 . 0 3 4 9 . 1 6

+ / -

1 . 4 3

1% Z110 8 . 0 7 5 . 4 0

+ / -

1 . 2 4 7 . 8 5 1 5 . 8 2

+ / -

1 . 6 8 6 . 0 8 5 8 . 1 9

+ / -

0 . 7 3

[ 0 1 4 3 ]

4 . Add P o l y g l y c e r y l - 2

d i p o l y h y d r o x y s t e a r a t e t o [ 0 1 6 3 ] The e s u l t s i n T a b l e 5

d e m o n s t r a t e

t h a t Z i n c o x i d e

t h e mixture a t 90-105o

C .

M i x a t 100-300 rpm u n t i l

h o m o g e n e o u s .

[ 0 1 4 4 ]

5 .

AlloW m i x t u r e t o c o o l t o 8 5 - 9 5 C and add

P o l y g l y c e r y l - 3 d i i s o s t e a r a t e . Mix t 1 0 0 - 3 0 0 rpm

n t i l

h o m o g e n e o u s .

[ 0 1 4 5 ]

6 . A d d a n o l i n BHT t

8 5 - 9 5 0 C .

M i x t 200-500

rpm

n t i l

h o m o g e n e o u s .

[ 0 1 4 6 ]

7 .

A d d i n c Oxide t 8 5 - 9 5 0 C . M i x t 200-500

rpm n t i l

h o m o g e n e o u s .

[ 0 1 4 7 ]

8 . HomogeniZe

m i x t u r e a t 5 0 0 0 - 6 0 0 0

rpm

o r 5

m i n u t e s .

[ 0 1 4 8 ]

9 . Remove i x t u r e

f r o m

h o m o g e n i Z e r

a n d k e e p

a t

85-900 C .

[ 0 1 4 9 ]

W a t e r

P h a s e

[ 0 1 5 0 ]

1 0 .

I n a s e c o n d b e a k e r , combine

Water 1

and

C i t r i c A c i d a t 4 0 - 5 0 0 C . M i x

t

100-300 rpm

n t i l homo

g e n e o u s .

[ 0 1 5 1 ]

1 1 .

I n

a t h i r d b e a k e r , combine Water and i s o

dium EDTA a t 40-500

C .

M i x

a t

100-300 rpm

u n t i l

h o m o g e n e o u s .

[ 0 1 5 2 ] 1 2 .

B r i n g m i x tu r e

t o 5 0 - 6 0 0 C .

a n d

a d d

Magne

sium S u l f a t e

H e p t a h y d r a t e . M i x t 1 0 0 - 3 0 0 rpm

n t i l

h o m o g e n e o u s .

[ 0 1 5 3 ] 13.Add h e n o x y e t h a n o l a t

5 0 - 6 0 0

C .

M i x

t 1 0 0

3 0 0

rpm

n t i l h o m o g e n e o u s .

h a s m i n i m a l

p r o t e a s e

a c t i v i t y , W h i l e t h e

m i x t u r e s

o f Z i n c

o x i d e a n d

c e t i c

a c i d h a v e t r y p s i n i n h i b i t o r y a c t i v i t y , W h i c h

i s

d o s e - d e p e n d e n t . T h i s s u g g e s t s t h a t t h e c o m b i n a t i on o f Z i n c

o x i d e , a knoWn

a g e n t

u s e d

t o r e l i e v e d i a p e r

r a s h , W i t h l o W

c o n c e n t r a t i o n s o f a c e t i c a c i d , c o u l d h a v e a s y n e r g i s t i c

e f f e c t

b y a c t i v e l y i n h i b i t i n g t h e p r o g r e s s i o n o f h e

r a s h .

TABLE

T r y p s i n i n h i b i t o r y a c t i v i t y

o f o r m u l a t i o n s

( p r e p a r a t i o n s

1 and

2 as de?ned a b o v e ) c o n t a i n i n g Z i n c o x i d e

and

a c e t i c a c i d ( t e s t e d

a t a 1 : 1 0 d i l u t i o n ) :

P r e p a r a t i o n - 1 P r e p a r a t i o n - 2

I n g r e d i e n t s

% r y p s i n i n h i b i t i o n % r y p s i n i n h i b i t i o n

ZnO, no a c e t i c a c i d 2 4 . 3 5 + / — 1 . 3 4 2 3 . 6 8 + / — 2 . 4 3

ZnO, 0.1 a c e t i c

a c i d

2 9 . 4 3 + / —

1 . 4 7

1 8 . 6 8 + / — 3 . 6 2

ZnO, 0.2 a c e t i c a c i d

7 0 . 8 7

+ / — 0 . 9 2

3 2 . 7 0

+ / — 6 . 0 9

ZnO,

0. 3 a c e t i c

a c i d 2 8 . 5 3 + / — 3 . 1 2

ZnO, 0. 4 a c e t i c a c i d

3 9 . 4 0 + / — 9 . 0 9

ZnO,

0. 5

a c e t i c a c i d 2 4 . 4 0 + / — 4 . 6 8

ZnO,

0.6 a c e t i c a c i d 5 0 . 1 4 + / — 4 . 2 5 3 9 . 1 3 + / — 1 3 . 7 5

[ 0 1 6 4 ] The e s u l t s i n T a b l e

6

s u g g e s t

t h a t a

p o r t i o n o f t h e

Z i n c o x i d e i s c o n v e r t e d t o Z i n c i o n s t h r o u g h t h e a d d i t i o n o f a

s m a l l amount

o f

a c i d i n t o t h e f o r m u l a , and t h a t i t i s

t h e

Z i n c

i o n s t h a t

a r e

a c t i v e

i n i n hi b i t i n g

t r y p s i n a c t i v i t y

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US 2013/0004580

A 1

TABLE

J a n .

3 ,

2 0 1 3

T r y p s i n

i n h i b i t o r y a c t i v i t y o f f o r m u l a t i o n c o n t a i n i n g ( 1 )

Z i n c o x i d e

a n d n o c i t r i c a c i d

( p r e p a r a t i o n

3 ) ;

re

aration

5 .

Zinc

oxide and

0 . 1

i t r i c

acid

r e a r a t i o n 4 3 Z i n c o x i d e and 0. 5 i t r i c a c i d

F o r m u l a t i o n

C u r r e n t l y

a v a i l a b l e

p r o d u c t

P r e p a r a t i o n

3

P r e p a r a t i o n

4

P r e p a r a t i o n 5

( B a b y P a s t e 5 0 * )

I n g r e d i e n t s B a b y d i a p e r r a s h B a b y d i a p e r r a s h B a b y d i a p e r r a s h F o r m u l a t i o n c o n t a i n s

cream base

with

cream base

with

cream base with 25% ZnO

and

an

13% ZnO 1 3 ZnO

and 0 .1

1 3 ZnO

and 0. 5 unknow n amount of

c i t r i c

a c i d

c i t r i c

a c i d a c e t i c

a c i d ,

pH

%

r y p s i n

i n h i b i t o r y

2 0 . 0 2 + / —

6 . 7 1

5 0 . 7 2 + / — 1 6 . 7 4 8 6 . 0 9 + / — 3 . 6 4 2 9 . 0 6 + / — 2 . 3 4

a c t i v i t y ( t e s t e d a t

1 : 5

d i l u t i o n )

* M e n d e l s o n P h a r m a c e u t i c a l s LLC, o u n t a i n s i d e , N J C o n t a i n s 2 5 i n c o x i d e ; w h i t e p e t r o l a t u m ; a n d a c e t i c a c i d f r o m v i n e g a r .

[ 0 1 6 5 ] I n a s e p a r a t e s t u d y , t h e a d d i t i o n

o f

0.5 i t r i c a c i d

( p r e p a r a t i o n - 6 ) o r 0.5 Z n C l 2 ( p r e p a r a t i o n - 7 ) t o t h e

p r e

made

b a b y d i a p e r r a s h

c r e a m

b a s e

b y

s i m p l e m i x i n g

was

t e s t e d a n d c o m p a r e d

w i t h a c o m m e r c i a l l y - a v a i l a b l e

p r o d u c t

f o r d i a p e r r a s h a f t e r a d d i n g 0 . 5 %

c i t r i c

a c i d ( p r e p a r a t i o n - 8 )

a n d s i m p l e

m i x i n g .

The e s u l t s a r e

shown

n

T a b l e

8 .

t r y p s i n i n h i b i t o r y

a c t i v i t y

b e c a u s e

o f

i n t e r f e r e n c e .

I n a d d i

t i o n , p r e p a r a t i o n 1 (10 ZnO

a n d

0 . 2 % a c e t i c a c i d ) , w h i c h

c o n t a i n s s i m i l a r i n g r e d i e n t s a s

Baby

P a s t e

5 . 0 ,

s h o w e d

s i g

n i ? c a n t l y h i g h e r a c t i v i t y ( 7 1 % , T a b l e 6 ) t h a n t h e l a t t e r ( 2 9 % ,

T a b l e 7 ) . T h e r e f o r e , w e o n c l u d e t h a t t h e Baby a s t e 5 . 0

a n d

t h e baby

d i a p e r r a s h

cream

b a s e

( p r e p a r a t i o n - 1 ) h a v e

n o

TABLE

C u r r e n t l y

a v a i l a b l e

p r o d u c t

( B a b y

F o r m u l a t i o n P r e p a p r a t i o n - 3 P r e p a r a t i o n - 4 P r e p a r a t i o n - 5

P a s t e

5 0 * ) P r e p a r a t i o n - 6 P r e p a r a t i o n - 7 P r e p a r a t i o n - 8

I n g r e d i e n t s b a b y d i a p e r b a b y d i a p e r b a b y d i a p e r f o r m u l a t i o n A d d i n g 0 . 5 %

A d d i n g

0 . 5 % A d d i n g 0 . 5 %

r a s h

cream

r a s h cream

r a s h

cream c o n t a i n s c i t r i c a c i d t o

ZnCl2

o p r e - c i t r i c a c i d t o

b a s e

(13

b a s e

w i t h

13 b a s e

w i t h

1 3

2 5 ZnO n d p r e - m a d e b a b y made a b y Baby a s t e 5 . 0

ZnO)

ZnO

a n d 0.1

ZnO n d

0.5

a n

unknown d i a p e r r a s h d i a p e r r a s h

c i t r i c a c i d c i t r i c a c i d amount of cream base cream base

a c e t i c a c i d ,

pH ( p r e p a r a t i o n 3 ,

( p r e p a r a t i o n

3 ,

13

ZnO)

13

1 1 0 )

r y p s i n 2 0 . 0 2 + / — 6 . 7 1 5 0 . 7 2 + / — 1 6 . 7 4 8 6 . 0 9 + / — 3 . 6 4 2 9 . 0 6 + / — 2 . 3 4 8 2 . 8 4 + / —

0 . 7 9

8 8 . 5 9 + / —

0 . 6 7

3 0 . 3 7 + / —

2 . 9 3

i n h i b i t o r y

a c t i v i t y ( t e s t e d

a t 1 : 5 d i l u t i o n )

* M e n d e l s o n

P h a r m a c e u t i c a l s

LLC, o u n t a i n s i d e , N J C o n t a i n s 2 5 i n c o x i d e ;

w h i t e

p e t r o l a t u m ; a n d a c e t i c a c i d f r o m v i n e g a r .

[ 0 1 6 6 ]

For

t h e above

s t u d y ,

t h e

r e s u l t s a r e an

a v e r a g e

of

t h r e e i n d e p e n d e n t e x p e r i m e n t s , w h e r e i n t h e

s a m p l e s ,

w h i c h

were i l u t e d a t 1 : 1 0

r a t i o

i n

1 > < P B S

s o l u t i o n s c o n t a i n i n g 10 ,

20% r 40%

t h a n o l ,

were

e s t e d

i n t r i p l i c a t e s . The

e s u l t s a r e

from

an

average of l l c o n d i t i o n s at t h e

same

d i l u t i o n .

[ 0 1 6 7 ]

F o r t h e c u r r e n t l y

a v a i l a b l e

p r o d u c t

( i . e . , B a b y P a s t e

5 . 0 ,

M e n d e l s o n

P h a r m a c e u t i c a l s

L L C ,

M o u n t a i n s i d e , N . J . ) ,

t h e r e s u l t s a r e

an

a v e r a g e

of

s i n g l e s t u d y :

sample

d i l u t e d a t

1 : 1 0 r a t i o , i n 1 > < P B S s o l u t i o n s c o n t a i n i n g

10 ,

2 0

r 4 0

e t h a n o l , was

e s t e d

i n t r i p l i c a t e .

The

e s u l t s

a r e from an a v e r

a g e

o f

l l

c o n d i t i o n s

a t

t h e

same i l u t i o n . The

Baby a s t e 5 . 0

( c o n t a i n i n g

25

n O )

a n d b a b y

d i a p e r r a s h c r e a m ( p r e p a r a

t i o n - 3 , c o n t a i n i n g 13

ZnO)

a v e

p l a c e b o

e f f e c t

i n t e r m s

o f

l o w

t r y p s i n

i n h i b i t o r y a c t i v i t y ( 2 9 . 0 6 % a n d 2 0. 0 2 % ,

r e s p e c

t i v e l y ) .

T h i s

e f f e c t

i s

most l i k e l y

d u e

t o t h e i n t e r f e r e n c e o f

s o l u b l e

o r

i n s o l u b l e

f o r m u l a t i o n

i n g r e d i e n t s

( e . g . ,

ZnO) i t h

t h e

a s s a y , and t h e r e f o r e , i t i s b e l i e v e d t h a t i t

i s

t h e

r e s u l t o f

a s s a y c o n d i t i o n s

a n d

n o t o f

i n h i b i t o r y

a c t i v i t y . P r e p a r a t i o n - 4

and 5

with 0 . 1 and 0 . 5

a c e t i c

a c i d showed a c i d

concen

t r a t i o n - d e p e n d e n t

i n c r e a s e i n t r y p s i n i n h i b i t o r y

a c t i v i t y

( 5 0 .

7 2

n d

8 6 . 0 9 % , r e s p e c t i v e l y ) .

T h e

p l a c e b o s o f t h e s e

p r e p a

r a t i o n s , which d i d n o t c o n t a i n

t h e a c e t i c a c i d ,

a l s o had l o w

s i g n i ? c a n t

t r y p s i n

i n h i b i t o r y a c t i v i t y ,

a n d

t h a t i t i s

t h e

a d d i

t i o n

o f c i t r i c

a c i d t o t h e

baby

d i a p e r r a s h

cream b a s e

which

l e a d s

t o t h e

r e l e a s e o f

Z i n c

i o n and e s u l t s i n i n c r e a s e d t r y p s i n

i n h i b i t o r y

a c t i v i t y .

[ 0 1

6 8 ]

The

o r e g o i n g e x a m p l e s a r e n o t i n t e n d e d t o

l i m i t

t h e

s c o p e o f t h e

p r e s e n t i n v e n t i o n , which

s

s e t o u t

i n t h e f o l l o w

i n g c l a i m s .

I n p a r t i c u l a r ,

v a r i o u s e q u i v a l e n t s a n d

s u b s t i t u t i o n s

w i l l b e r e c o g n i z e d by

t h o s e

s k i l l e d i n t h e

a r t i n v i e w

o f t h e

f o r e g o i n g d i s c l o s u r e

and

h e s e

a r e

c o n t e m p l a t e d

t o

b e

w i t h i n

t h e s c o p e o f t h e i n v e n t i o n .

1 . A

o m p o s i t i o n

f o r t r e a t i n g a n d / o r p r e v e n t i n g e n z y m a t i c

d e r m a t i t i s , c o m p r i s i n g

i )

a d i v a l e n t

c a t i o n

o f m a g n e s i u m ,

i i )

t a l c and

i i ) a c a r r i e r .

2 .

The

o m p o s i t i o n

o f c l a i m

1 ,

w h e r e i n t h e

d i v a l e n t

c a t i o n

o f magnesium

s

p r e s e n t

i n an amount e l e c t e d from h e g r o u p

c o n s i s t i n g

o f

f r o m

a b o u t 0.001 o a b o u t 50 ,

from

a b o u t

0.001

to

a b o u t 30 , from a b o u t 0.01

to

a b o u t 10 , and

from a b o u t 0.1

t o

a b o u t 5 . 0 % .

3 .

The

o m p o s i t i o n

o f c l a i m

2 ,

w h e r e i n t h e

d i v a l e n t

c a t i o n

i s p r e s e n t i n an amount of from a b o u t

0.001

t o

a b o u t 30%

d i v a l e n t c a t i o n .

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US 2013/0004580

A 1

4 .

The

o m p o s i t i o n o f

c l a i m

3 , w h e r e i n t h e d i v a l e n t c a t i o n

i s

p r e s e n t

i n

an

amount of

from

a b o u t 0 . 0 1

t o

a b o u t

1 0

d i v a l e n t

c a t i o n .

5 .

The

o m p o s i t i o n

o f c l a i m

4 ,

W h e r e i n t h e d i v a l e n t c a t i o n

i s

p r e s e n t

i n

an

amount

of

from

a b o u t

0.1

to

a b o u t 5.0 .

6 . The

o m p o s i t i o n

o f c l a i m

5 ,

W h e r e i n t h e

d i v a l e n t

c a t i o n

i s p r e s e n t i n an amount of from a b o u t 0 . 1 to

a b o u t

1.0

d i v a l e n t c a t i o n .

7 .

The

o m p o s i t i o n o f

c l a i m

1 ,

W h e r e i n

t h e d i v a l e n t

c a t i o n

o f magnesium s

p r e s e n t

a s a d i v a l e n t

c a t i o n /

a n i o n

p a i r .

8 .

The

o m p o s i t i o n o f c l a i m

7 ,

W h e r e i n t h e d i v a l e n t c a t i o n /

a n i o n p a i r i s s e l e c t e d f r o m t h e g r o u p c o n s i s t i n g o f magne

s i u m

c h l o r i d e , m a g n e s i u m c e t a t e , m a g n e s i u m g l u c o n a t e

a n d

m a g n e s i u m l a c t a t e .

9 . The c o m p o s i t io n o f c l a i m 1 , W h e r e i n t h e c a r r i e r i s

s e l e c t e d f r o m t h e g r o u p c o n s i s t i n g o f

o t i o n ,

c r e a m , g e l , s t i c k ,

s p r a y , o i n t m e n t , c l e a n s i n g l i q u i d W a s h , s o l i d b a r , p a s t e , poW

d e r ,

m o u s s e , W i p e ,

p a t c h , Wound r e s s i n g ,

a d h e s i v e

b a n d a g e ,

h y d r o g e l , ? l m a n d d i a p e r .

1 0 .

T h e

c o m p o s i t i o n

o f

c l a i m 1 , W h e r e i n

s a i d

c o m p o s i t i o n

h a s

a pH of from a b o u t 4 . 5 t o a b o u t 8 .

1 1 . T h e

c o m p o s i t i o n o f

c l a i m 1 0 , W h e r e i n s a i d c o m p o s i

t i o n

h a s

a pH of from

a b o u t

5 t o

a b o u t

6 .

1 2 . T h e

c o m p o s i t i o n o f

c l a i m 1 1 ,

W h e r e i n

s a i d c o m p o s i

t i o n

h a s

a pH of from

a b o u t

5 t o

a b o u t

5 . 5 .

J a n .

3 ,

2 0 1 3

1 3 .

T h e

c o m p o s i t i o n o f

c l a i m

1 2 ,

W h e r e i n s a i d c o m p o s i

t i o n

h a s a

pH f a b o u t 5 . 5 .

1 4 .

The

c o m p o s i t i o n

o f c l a i m

1 , f u r t h e r c o m p r i s i n g

a n

a g e n t

s e l e c t e d f r o m

t h e g r o u p c o n s i s t i n g o f

k e t o c o n a Z o l e ,

m i c o n a Z o l e ,

e l u b i o l , a l l a n t o i n ,

c a l a m i n e , d i m e t h i c o n e ,

k a o l i n , p e t r o l a t u m , W h i t e p e t r o l a t u m , c o d

l i v e r

o i l , l a n o l i n ,

m i n e r a l o i l ,

a n d t o p i c a l s t a r c h .

1 5 .

The c o m p o s i t i o n o f c l a i m 1 , W h e r e i n t h e enzyme s a

p r o t e a s e .

1 6 .

The c o m p o s i t i o n o f

c l a i m 1 5 , W h e r e i n

t h e

p r o t e a s e i s

t r y p s i n .

1 7 . A e t h o d f o r t r e a t i n g a n d / o r p r e v e n t i n g e n z y m a t i c

d e r m a t i t i s ,

c o m p r i s i n g

a d m i n i s t e r i n g

t h e c o m p o s i t i o n o f

c l a i m 1 .

1 8 . The method o f c l a i m

1 7 ,

Wherein t h e enZyme i s a

p r o t e a s e .

1 9 .

The

method o f c l a i m 1 8 , W h e r e i n t h e p r o t e a s e i s

t r y p s i n .

2 0 . The

method

o f

c l a i m

1 9 ,

W h e r e i n

t h e c o m p o s i t i o n

e x h i b i t s a t r y p s i n i n h i b i t o r y a c t i v i t y s e l e c t e d f r o m t h e g r o u p

c o n s i s t i n g

o f t

l e a s t a b o u t 15 , t l e a s t a b o u t

25 ,

t l e a s t

a b o u t

50 ,

t

l e a s t

a b o u t

60 ,

t

l e a s t

a b o u t 70 , t

l e a s t

a b o u t

80% and t l e a s t

a b o u t

90 .