Massachusetts Institute of Technology

University of Texas System

University of Pennsylvania

The Broad Institute, Inc.

Other

Yale University

Monty Krieger

Susan Laurene Acton

Attilio Rigotti

Benito Munoz

Sivaraman Dandapani

Partha Pratim Nag

Karen Kozarsky

Miao Yu

Chris Dockendorff

Willmen Youngsaye

Patrick Faloon

Thomas Nieland

Helena E Miettinen

Timothy A Lewis

Jay M Edelberg

Andrew Germain

Helen H Hobbs

Anne Braun

Bernardo Trigatti

Alan M Pearson

Kosuke Tsukamoto

Tatsuhiko Kodama

Keith A Joiner

Katherine Tallman Landschulz

Songwen Zhang

Sharon L Karackattu

Xiaoling Li

Leonard Guarente

This application describes compounds and methods that can inhibit Scavenger receptor class B, type I (SR-BI) activity, which compounds and methods can used, for example, to mediate high-density lipoprotein (HDL) lipid uptake and treat hepatitis C viral infections.

Thiazole-based inhibitors of scavenger receptor BI

This application describes compounds and methods that can inhibit Scavenger receptor class B, type I (SR-BI) activity, which compounds and methods can be used, for example, to mediate high-density lipoprotein (HDL) lipid uptake and treat hepatitis C viral infections. The compounds have the formula: wherein Rand Rare independently H, halogen, cyano, haloalkyl, haloalkyloxy or OMe; or Rand Rtogether are &#x2014;O&#x2014;CH&#x2014; or &#x2014;O&#x2014;CF&#x2014;O&#x2014;; Ris H, halogen, cyano, haloalkyl or haloalkyloxy; Ris Calkyl, Ccycloalkyl, Ccycloalkylmethyl or Ccycloheteroalkyl, wherein the heteroatom is N or O; Ris H or CH; Ris Calkyl or Ccycloalkyl; and A, B, D and E are each, independently, CH, N, O or S, wherein at least one of A, B, D and E is N, and another of A, B, D and E is N, O or S. 

Heterocycle-bisamide inhibitors of scavenger receptor BI

The invention provides methods for treating tissue damage associated with impaired blood flow using ApoD, or an active variant thereof, or an agent that increases the expression of ApoD. The invention also encompasses active variants of ApoD for use in the methods.

Methods for treating tissue damage associated with ischemia with apolipoprotein D

An animal model of coronary heart disease has been developed where myocardial infarct can be induced by altering the animal's diet. In all embodiments, this animal model is a result of reduced activity of scavenger receptor class BI (SR-BI) and apolipoprotein E (ApoE). In a preferred embodiment, the model is a result of crossbreeding two transgenic mouse lines: a knockout of SR-BI (SR-BI&#x2212;/&#x2212;) and an impaired ApoE expressor (hypoE). The impaired ApoE gene results in only 2-5% expression of ApoE and a reduction in cholesterol homeostasis. Resulting animals are predisposed to hypercholesterolemia but can live longer than a year on a normal low fat diet. Serum plasma levels can be significantly elevated by changing the animal's diet to one containing high levels of fat and cholesterol. Within a month on a high fat, high cholesterol diet, animals develop atherosclerosis and myocardial infarction occurs. Survival depends on the nature of the diet and the conditions of animal husbandry and can typically be around 20-30 days after administration of the modified diet depending on the specific conditions. Housing the animals alone or in groups significantly affects survival of these animals on a high fat diet. Analysis of B- and T-cell deficient SR-BI/ApoE/RAG2 triple knockout mice established that B- and T-lymphocytes do not play a key role in the pathophysiology of the SR-BI ApoE dKO model of human disease. These animal models can be used to study mechanisms and progression of CHD as a function of diet, treatment with drugs to be screened for efficacy or undesirable side effects, and social environmental effects.

Inducible heart attack animal model

Transgenic animals that do not express functional SR-BI and ApoE develop severe atherosclerosis, by age four weeks in transgenic mice. Moreover, these animals exhibit progressive heart dysfunction by as early as age four weeks, and die by age nine weeks. This animal model has now been demonstrated to be useful as a screen for compounds which alleviate the symptoms of atherosclerosis and heart disease. Animals (Apo E&#x2212;/&#x2212; SR-BI+/&#x2212;) were fed PROBUCOL beginning at the time of mating. Offspring are weaned at three weeks and fed PROBUCOL. In contrast to animals (Apo E&#x2212;/&#x2212; SR-BI&#x2212;/&#x2212;) not fed PROBUCOL, 50% of whom are dead at six weeks, all animals (Apo E&#x2212;/&#x2212; SR-BI&#x2212;/&#x2212;) on PROBUCOL have a normal phenotype (MRI of heart function, ECG, echocardiogram, histology) at six weeks. At seven to eight months, there is evidence of atherosclerosis and some myocardial infarction. This demonstrates that the compound has a preventative action. Animals who are taken off of the PROBUCOL all die within ten to twelve weeks. In another study, the majority of animals whose parents were not fed PROBUCOL, but who received the PROBUCOL beginning at about five weeks of age, survived for a few months, demonstrating that the compound also has a therapeutic benefit.

Screening of compounds for treatment of atherosclerosis and heart attack

SR-BI is present at relatively high levels on the membranes of hepatocytes and steroidogenic tissues, including the adrenal gland, testes, and ovaries, where it mediates the uptake and transport of cholesteryl ester from high density lipoproteins. It has been demonstrated that transgenic animals which do not produce SR-BI are healthy, with the exception that the females are infertile. SR-BI KO females have abnormal HDLs, ovulate dysfunctional oocytes and are infertile. Surgical, genetic and pharmacologic methods were used to show that the fertility of SR-BI KO females (or their transplanted oocytes) can be restored in the absence of ovarian and/or extraovarian SR-BI expression by manipulations that modify the structure, composition and/or abundance of their abnormal plasma lipoproteins. These manipulations included inactivation of the apolipoprotein A-I gene and administration of the cholesterol-lowering drug PROBUCOL&#x2122;. In the absence of treatment, female animals which do not express SR-BI have dramatically reduced levels of offspring, even though they are otherwise healthy and the males normal. Studies demonstrate that they do not produce viable eggs and have a defect involving implantation of normal eggs.

Lipid-altering compositions for the treatment of infertility

Two distinct scavenger receptor type proteins having high affinity for modified lipoproteins and other ligands have been isolated, characterized and cloned. HaSR-BI, an AcLDL and LDL binding scavenger receptor, which is distinct from the type I and type II macrophage scavenger receptors, has been isolated and characterized and DNA encoding the receptor cloned from a variant of Chinese Hamster Ovary Cells, designated Var-261. dSR-CI, a non-mammalian AcLDL binding scavenger receptor having high ligand affinity and broad specificity, was isolated from. The isolated receptors are useful in screening for drugs that inhibit uptake of cholesterol in endothelial or adipose cells or macrophages, respectively. They are also useful as probes for the isolation of other lipoprotein receptors and in research the roles of these receptors.

Class BI and CI scavenger receptors

Methods for regulation of lipid and cholesterol uptake are described which are based on regulation of the expression or function of the SR-BI HDL receptor. The examples demonstrate that estrogen dramatically downregulates SR-BI under conditions of tremendous upregulation of the LDL-receptor. The examples also demonstrate the upregulation of SR-BI in rat adrenal membranes and other non-placental steroidogenic tissues from animals treated with estrogen, but not in other non-placental non-steroidogenic tissues, including lung, liver, and skin. Examples further demonstrate the uptake of fluorescently labeled HDL into the liver cells of animal, which does not occur when the animals are treated with estrogen. Examples also demonstrate the in vivo effects of SR-BI expression on HDL metabolism, in mice transiently overexpressing hepatic SR-BI following recombinant adenovirus infection. Overexpression of the SR-BI in the hepatic tissue caused a dramatic decrease in cholesterol blood levels. These results demonstrate that modulation of SR-BI levels, either directly or indirectly, can be used to modulate levels of cholesterol in the blood.

Methods for modulation of cholesterol transport

Transgenic animals that do not express functional SR-BI and ApoE develop severe atherosclerosis, by age four weeks in transgenic mice. Moreover, these animals exhibit progressive heart block by age four weeks, and die by age nine weeks. Pathology shows extensive fibrosis of the heart and occlusion of coronary arteries. The occlusion appears to be due to clotting, since fat deposition is in the walls. These animals are good models for the following diseases, and for screening of drugs useful in the treatment and/or prevention of these disorders: cardiac fibrosis, myocardial infarction, defects in electrical conductance, atherosclerosis, unstable plaque, and stroke. In contrast to other known models for atherosclerosis, these animals do not have to be fed extreme diets for long periods before developing atherosclerosis. No other known model for heart attacks and stroke is known.

SR-BI and ApoE knockout animals and use thereof as models for atherosclerosis and heart attack

Two distinct scavenger receptor type proteins having high affinity for modified lipoproteins and other ligands have been isolated, characterized and cloned. HaSR-BI, an AcLDL and LDL binding scavenger receptor, which is distinct from the type I and type II macrophage scavenger receptors, has been isolated and characterized and DNA encoding the receptor cloned from a variant of Chinese Hamster Ovary Cells, designated Var-261. dSR-CI, a non-mammalian AcLDL binding scavenger receptor having high ligand affinity and broad specificity, was isolated from . The isolated receptors are useful in screening for drugs that inhibit uptake of cholesterol in endothelial or adipose cells or macrophages, respectively. They are also useful as probes for the isolation of other lipoprotein receptors and in research the roles of these receptors.

Two distinct scavenger receptor type proteins having high affinity for modified lipoproteins and other ligands have been isolated, characterized and cloned. HaSR-BI, an AcLDL and LDL binding scavenger receptor, which is distinct from the type I and type II macrophage scavenger receptors, has been isolated and characterized and DNA encoding the receptor cloned from a variant of Chinese Hamster Ovary Cells, designated Var-261. dSR-CI, a non-mammalian AcLDL binding scavenger receptor having high ligand affinity and broad specificity, was isolated from The isolated receptors are useful in screening for drugs that inhibit uptake of cholesterol in endothelial or adipose cells or macrophages, respectively. They are also useful as probes for the isolation of other lipoprotein receptors and in research the roles of these receptors.

Methods for regulation of lipid and cholesterol uptake are described which are based on regulation of the expression or function of the SR-BI HDL receptor. The examples demonstrate that estrogen dramatically downregulates SR-BI under conditions of tremendous upregulation of the LDL-receptor. The examples also demonstrate the upregulation of SR-BI in rat adrenal membranes and other non-placental steroidogenic tissues from animals treated with estrogen, but not in other non-placental non-steroidogenic tissues, including lung, liver, and skin. Examples further demonstrate the uptake of fluorescently labeled HDL into the liver cells of animal, which does not occur when the animals are treated with estrogen.

Methods for modulation of lipid uptake

Macrophage scavenger receptor protein or active fragments thereof bind specifically to the lipoteichoic acid residues on the Gram-positive bacterial cell wall. This protein or active fragments thereof can be used in a variety of methods including methods to specifically purify lipoteichoic acid, to preferentially label or detect lipoteichoic acid or lipoteichoic acid-containing compounds or cells, and to treat patients having Gram-positive bacterial infections such as septicemia and associated pathophysiological states such as septic shock.

Method for treating gram positive septicemia

A substantially pure receptor protein capable of binding acetylated low density lipoprotein and oxidized low density lipoprotein is disclosed herein. This protein is characterized by having a molecular weight of about 220,000 daltons, and an affinity for oxidized low density lipoprotein and acetylated low density lipoprotein. Further, it is an integral membrane protein which includes a collagen domain. Proteins having an affinity for the receptor protein as well as DNA sequences encoding at least a portion of the receptor protein are also disclosed herein. In addition, devices for purification purposes and methods for detecting atherosclerotic plaques are described, both of which utilize the receptor protein or binding proteins thereto.

Growing community of inventors

Needham, MA, United States of America

Monty Krieger