vitamin d current outlook, design and synthesis of active analogs

Vitamin D: Current Outlook, Design and Synthesis of Active Analogs

UNIVERSIDAD DE SANTIAGO DE COMPOSTELADepartamento de Química OrgánicaSantiago de Compostela E-15782, EspañaAntonio Mouriño: [email protected]

Development of Active vitamin D Analogues

UNIVERSIDAD DE SANTIAGO DE COMPOSTELADepartamento de Química OrgánicaSantiago de Compostela E-15782, EspañaAntonio Mouriño: [email protected]

Birmingham. DECIDE. Nov 14, 2013miércoles 27 de noviembre de 2013

miércoles 27 de noviembre de 2013

Alimentos

VIT-D3

PRE-D3

7-Deshidrocolesterol25-D3

1,25-D3

Forma Activa

80%20%

Active form 1,25D

Food

7-Dehydrocholesterol


Vitamin D Metabolism

7-Dehydrocholesterol Previtamin D Cisoid form

Transoid form

Vitamin D

Kidney Liver

3

3

1,25D

(calcitriol)


.

.

HO

OHHO

H

H

1α,25-(OH)2-D3

ICABCM

.

.

BIOLOGICALRESPONSES


.

.

HO

OHHO

H

H

1α,25-(OH)2-D3

ICABCM

DIFFERENTIATION PROLIFERATION

Miyaura et alBiochem. Biophys. Res.Commun.1981, 102, 937

.

.

BIOLOGICALRESPONSES


.

.

HO

OHHO

H

H

1α,25-(OH)2-D3

ICABCM

DIFFERENTIATION PROLIFERATION

Miyaura et alBiochem. Biophys. Res.Commun.1981, 102, 937

BONE DISEASES(OSTEOPOROSIS)

IMMUNOLOGY(TRANSPLANTATION)

PSORIASIS

CANCER

.

.


DT-71


HO

H

H

OH

OH

HO

H

H

OH

OH

HO

H

H

OH

OH

HO

H

H

OH

OH

HO

H

H

OH

OH


HO

H

H

OH

HO


Cytoplasm

Nucleus

Mode of action of 1α,25-(OH)2-D3


Cytoplam

Nucleus

Mode of action of 1,25D


Cytoplam

Nucleus


RXR


Target gene

Cytoplam

Nucleus


RXR

coA

m-RNA


Target gene

Cytoplam

Nucleus


Proteins

RXR

coA

m-RNA


Vitamin D Nuclear Receptor (VDR)


Receptor Nuclear de la Vitamina D (VDR)

DNA



Shaffer PL, Gewirth DT.

EMBO J. 2002, 21, 2242-2252

DNA



1α,25-(OH)2-D3



1α,25-(OH)2-D3

Moras D. y col.Mol. Cell 2000, 5, 173-179


Ligand Binding Domain

H12

H11

H1

H2

H3

H4

H5

H6

H8H9


2.81

2.82

Ser278

2.91

2.83

2.832.86

Ser237

Ligand Binding Domain


Prof Roger Bouillon and Mieke VerstuyfLeuven University

Transactivation, Calcemic Effects, Cell Differentiation-Proliferation

Prof. Román Pérez FernándezFaculty of Medicine-USCSantiago Compostela

Transactivation, Cell Diffentiation-Proliferation(colon cancer cell lines)

Prof Alberto MuñózInstitute of Biomedical Investigations CSIC-Madrid

Transactivation, Calcemic Effects, Cell Differentiation-Proliferation

Prof. Dino Moras and Natacha RochelIGBMC-Illkirch-France Structural Studies

COLLABORATIONS


Superposition of Diyne (Docking Conformation)

with 1,25D (Crystallographic Structure)


Drop containing crystals of Moras mutant VDR LBD complexed to 4cHHOHHOOH4c

Diyne-LBD(VDR)-

Complex HHO

HO

OH

VDR

Prof. Dino Moras (GBMC-Illkirch-Francia)


N Rochel, S Hourai, X Pérez, A Rumbo, A Mouriño

Arch. Biochem. Biophys. 2007, 460, 172-176

Crystallographic Structue of the Diyne superimposed with 1,25D


HO

H

H

OH

OH

12

HO

H

H

OH

OH

12


1,25D with a Me Group at C-12


?

Exploration of side

chains at C-12

Docking

12

Design, Syntheses and Biological Assays of

Vitamin Analogues with Side Chains at C-12.

J. Med. Chem. 2006, 49, 1509-1516

OH

H

OHHO

H

OHHO

OH

n

n = 1, 2, 3

1,25D Analogs with Side Chains at C-12 1,25D Analogs with Side Chains at C-12


(104%) (113%) (112%)


in silico LBD(VDR) 100 104 113 112

Affinity for hVDR 100 17 22 13

Transactivation (10-9 M) 100 40 36 57

Ca Effects 100 0 18 1

J. Med. Chem. 2012, 55, 8642


Analogs of 1,25D with Side Chains at C-18


.


DS-A and DS-B induced expression of the protein E-cadherin comparably to “1,25”

E-cadherin

VDR

-actin

Time (h)1α,25-D 3

Vehicle

Protein

1.0 1.6 6.3 1.7 4.6 0.8 1.5

DS-A DS-B 8 48 8 48 8 4848

1.0 3.4 2.1 2.4 1.6 0.9 0.8

Vehicle DS-A

48 h

DS-B 1α,25-D3

A

B

Figure X

Texto

DS-A y DS-B Induce cell differentiation in colon

cancer cell lines (SW-480-ADH) similarly to 1,25D


5

10

15

20

25

30

VD

RE

act

ivat

ion

(fold

incr

ease

)

010- 710- 810- 9 10- 710- 810- 9 10- 710- 810- 9

DS-A DS-B 1α,25-D3

Figure Y

DS-A and DS-B activate VDR transactivation in human colon cancer cells (SW480-ADH).


0

10

20

30

40

50

Cel

l pro

lifer

atio

n (%

inhi

bitio

n)

DS-A DS-B 1α,25-D3

Figure Z

DS-A and DS-B inhibit cell proliferation (50% vs 1,25)


Structures of Superagonists MC1288 y KH1060

Superagonists: Potent inducers of cell

differentiation


Superimposition of the crystallographic

structures of 1,25D and superagonists

KH1060 y MC1288

Moras D et al.

PNAS 2001, 98, 5491-5496


Rational Design of a New Superagonist

New Superagonist?


54

A B


Isomer A induces transactivation 12 times better than

1,25D at 10-10 M, The isomer B behaves as 1,25D.

A

B1,25


58

NEW SUPERAGONIST?

29

DESIGN OF A NEW SUPERAGONIST

+ =

2-Me-1,25D BINDS 4 TIMES BETTER TO VDR THAN 1,25D

A-MeISOMER A


Interaction Energies with the

A-Me B-Me


TRANSACTIVATION ACTIVITY

A-Me is 13 times more potent than 1,25D and 4 times more potent than A at 10 M

A A-Me1,25D

-10


65

• Proliferación celular en HL-60 • Diferenciación celular en HL-60

Biological Activity Similar than 1,25D


66

Arg274

Gln277

Tyr147

Ser278

Ser237

Tyr143

W1

W2

W3

Superimpositionof the New Metabolite with 1,25D


Prof. Román Pérez-Fernández


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