阅读说明：本技术 一种平滑受体配体 (Smooth receptor ligand ) 是由 陶厚朝 赵飞 于 2018-09-13 设计创作，主要内容包括：本发明涉及生物技术领域,特别是涉及一种平滑受体配体。本发明提供一种平滑受体配体或其异构体前体药物、溶剂化物、药学上可接受的盐,所述平滑受体配体的结构式为：A——linker——B其中,A为胞外域配体结构,B为跨膜域配体结构；Linker为线性的、对平滑受体无活性的亚基。本发明所提供的针对平滑受体的新型双头小分子配体,结合平滑受体的晶体结构数据,在胞外域配体和跨膜域配体的适当位点引入连接体,得到全新的双头配体小分子,增强了配体和受体之间的相互作用以及配体的生物活性。(The invention relates to the field of biotechnology, in particular to a smooth receptor ligand. The invention provides a smooth receptor ligand or an isomer prodrug, solvate and pharmaceutically acceptable salt thereof, wherein the structural formula of the smooth receptor ligand is as follows: A-linker-B, wherein A is an extracellular domain ligand structure, and B is a transmembrane domain ligand structure; linker is a linear, smooth receptor inactive subunit. The novel double-headed small molecule ligand for the smooth receptor provided by the invention is combined with the crystal structure data of the smooth receptor, and a connector is introduced into a proper site of an ectodomain ligand and a transmembrane domain ligand to obtain a brand-new double-headed ligand small molecule, so that the interaction between the ligand and the receptor and the biological activity of the ligand are enhanced.)

1. A smoothing receptor ligand, or an isomer prodrug, solvate, pharmaceutically acceptable salt thereof, having the structural formula:

A-linker-B

wherein A is an ectodomain ligand structure, and B is a transmembrane domain ligand structure;

linker is a linear, smooth receptor inactive subunit.

2. The smoothing receptor ligand of claim 1, wherein the ectodomain ligand structure is a steroid structure.

3. The smoothing receptor ligand of claim 1, wherein a is selected from the group consisting of:

wherein E is selected from H, -OH;

f is selected from linear C1-C20 aliphatic groups containing or not containing heteroatoms selected from N, S, P, O;

g is selected from linear C1-C20 alkyl or acyl with or without a heteroatom selected from N, S, P, O.

4. The smoothing receptor ligand of claim 1, wherein a has a structure selected from the group consisting of 20- (S) -hydroxycholesterol, 22- (S) -azacholesterol, cyclopamine, cholesterol, lithocholic acid, and 20- (S) -hydroxy-24-alkynylcholesterol.

5. The smoothing receptor ligand of claim 1, wherein a is selected from the group consisting of:

6. the smoothing receptor ligand of claim 1, wherein the transmembrane domain ligand structure is an amide or imine-like structure containing multiple aliphatic rings, nitrogen-containing aliphatic rings, benzene rings, nitrogen-containing aromatic ring systems.

7. The smoothing receptor ligand of claim 1, wherein B is selected from the group consisting of:

wherein J is selected from substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, substituted or unsubstituted pyridyl, substituted or unsubstituted cycloalkyl, wherein the substituents are each independently selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, nitro, amino, hydroxy, alkoxy, cycloalkyl, aryl, heteroaryl;

l is selected from hydrogen, alkyl and substituent groups are selected from-NR 'R' substituted or unsubstituted cycloalkyl, wherein R 'and R' are independently selected from C1-C6 alkyl;

m is selected from substituents selected from F, Cl, Br, I, C1-C6 alkyl, C1-C6 alkoxy, -CF₃、-OCF₃Substituted or unsubstituted aryl or heteroaryl of alkylsulfonyl.

8. The smoothing receptor ligand of claim 1, wherein the structure of B is selected from the group consisting of SAG, LY2940680, GDC0449, LDE225, SANT1, SANT 2.

9. The smoothing receptor ligand of claim 1, wherein B is selected from the group consisting of:

10. the smoothing receptor ligand of claim 1, wherein the linker is a polyethylene glycol oligomer having a triazole structure.

11. The smoothing receptor ligand of claim 1, wherein the linker has the formula:

wherein m and n are independently selected from integers between 0 and 7.

12. The smoothing receptor ligand of claim 1, wherein the smoothing receptor ligand is selected from the group consisting of:

13. use of the smoothing receptor ligand of any one of claims 1-10, or an isomer, prodrug, solvate, pharmaceutically acceptable salt thereof, for the preparation of a smoothing receptor agonist, inhibitor or antagonist.

14. A pharmaceutical composition comprising a smoothing receptor ligand according to any one of claims 1-10, or an isomer, prodrug, solvate, or pharmaceutically acceptable salt thereof.

Technical Field

The invention relates to the field of biotechnology, in particular to a smooth receptor ligand.

Background

Smoothing receptors belong to the F family of G-protein-coupled receptors (GPCRs), whose dysfunction leads to birth defects. In the 50 s of the 20 th century, many born lambs were found to have only one eye in the western united states, and the first natural plant small molecule ligand Cyclopamine (Cyclopamine) for the smooth receptor was discovered through studies on this cyclopia. Functional regulation of smooth receptors is associated with the development of a range of cancers, and thus smooth receptors are important targets in drug development. Over the past few decades, a number of lead compound ligands have been discovered, with two small molecule drugs being marketed in 2015. However, resistance to drugs caused by the smooth receptor mutants rapidly develops, and further modification of existing drugs is urgently needed.

Disclosure of Invention

In view of the above-mentioned drawbacks of the prior art, it is an object of the present invention to provide a smoothing receptor ligand, which solves the problems of the prior art.

To achieve the above objects and other related objects, the present invention provides, in one aspect, a smoothing receptor ligand, or an isomer prodrug, solvate, or pharmaceutically acceptable salt thereof, wherein the smoothing receptor ligand has the structural formula:

A-linker-B

wherein A is an ectodomain ligand structure, and B is a transmembrane domain ligand structure;

linker is a linear, smooth receptor inactive subunit.

In some embodiments of the invention, the extracellular domain ligand structure is a steroid structure.

In some embodiments of the invention, a is selected from the group shown below:

wherein E is selected from H, -OH;

f is selected from linear C1-C20 aliphatic groups containing or not containing heteroatoms selected from N, S, P, O;

g is selected from linear C1-C20 alkyl or acyl with or without a heteroatom selected from N, S, P, O.

In some embodiments of the invention, A has a structure selected from the group consisting of 20- (S) -hydroxycholesterol, 22- (S) -azacholesterol, cyclopamine, cholesterol, lithocholic acid, and 20- (S) -hydroxy-24-alkynylcholesterol.

In some embodiments of the invention, a is selected from the group shown below:

in some embodiments of the invention, the transmembrane domain ligand structure may be an amide or imine-like structure containing multiple aliphatic rings, nitrogen-containing aliphatic rings, benzene rings, aza-aromatic ring systems.

In some embodiments of the invention, B is selected from the group shown below:

wherein J is selected from substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, substituted or unsubstituted pyridyl, substituted or unsubstituted cycloalkyl, wherein the substituents are each independently selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, nitro, amino, hydroxy, alkoxy, cycloalkyl, aryl, heteroaryl;

l is selected from hydrogen, alkyl and substituent groups are selected from-NR 'R' substituted or unsubstituted cycloalkyl, wherein R 'and R' are independently selected from C1-C6 alkyl;

m is selected from substituents selected from F, Cl, Br, I, C1-C6 alkyl, C1-C6 alkoxy, -CF₃、-OCF₃Substituted or unsubstituted aryl or heteroaryl of alkylsulfonyl.

In some embodiments of the invention, the structure of B is selected from SAG, LY2940680, GDC0449, LDE225, SANT1, SANT 2.

In some embodiments of the invention, B is selected from the group shown below:

in some embodiments of the invention, the linker is a polyethylene glycol oligomer having a triazole structure. In some embodiments of the invention, the linker has the following structural formula:

wherein m and n are independently selected from integers between 0 and 7.

In some embodiments of the invention, the smoothing receptor ligand is selected from the group consisting of the compounds shown below:

in another aspect, the invention provides the use of the smooth receptor ligand or its isomer, prodrug, solvate, pharmaceutically acceptable salt in the preparation of smooth receptor agonist, inhibitor or antagonist.

In another aspect, the invention provides a pharmaceutical composition comprising the smooth receptor ligand or its isomer, prodrug, solvate, pharmaceutically acceptable salt

Drawings

FIG. 1 is a schematic diagram showing the molecular orientation and spacing of extracellular domain ligands and transmembrane domain ligands confirmed by the crystal structure according to the present invention.

FIG. 2 shows a schematic representation of the simultaneous binding of two domains of the smooth receptor by the bipitch molecule of the present invention.

FIG. 3 shows a U-shaped curve of the activity of a series of double-headed molecules of the invention versus linker length.

Detailed Description

The inventor integrates the receptors of each binding domain of the smooth receptor together through a proper connector, can realize that the small molecule drug simultaneously binds two structural domains through the proximity effect, thereby enhancing the drug activity and realizing the design of the small molecule drug for avoiding drug resistance, and the invention is completed on the basis.

In one aspect, the present invention provides a smooth receptor ligand, or an isomer (e.g., enantiomer, diastereomer, geometric isomer, tautomer, rotamer, atropisomer, racemate, etc.), prodrug, solvate, or pharmaceutically acceptable salt thereof, having a formula:

A-linker-B

wherein A is an ectodomain ligand structure, and B is a transmembrane domain ligand structure;

linker is a linear, smooth receptor inactive subunit.

In the smoothing receptor ligands provided herein, a is an extracellular domain ligand structure, which generally refers to a molecular structure capable of recognizing and binding to the extracellular domain of a smoothing receptor, which may be specific. The ectodomain ligand structure may be an inhibitor structure, an antagonist structure, or an agonist structure. The ectodomain ligand structure may be a steroid structure, for example, a may be selected from the group shown below:

wherein E is selected from H (hydrogen atom), -OH (hydroxyl group); f is selected from C1-C20, C1-C12, C1-C6 and C1-C3 straight-chain aliphatic groups containing or not containing heteroatoms, and in the group F, the heteroatoms can be N, S, P, O; g is selected from C1-C20, C1-C12, C1-C6 and C1-C3 straight-chain alkyl or acyl containing or not containing heteroatoms, and in the group G, the heteroatoms can be N, S, P, O and the like. More specifically, the ectodomain ligand structure may be a group structure corresponding to, but not limited to, 20- (S) -hydroxycholesterol, 22- (S) -azacholesterol, cyclopamine, cholesterol, lithocholic acid, 20- (S) -hydroxy-24-alkynyl cholesterol, and other steroids, and the like. In other embodiments of the present invention, a may be selected from the group shown below:

in the ligand of the smooth receptor provided by the invention, B is a transmembrane domain ligand structure, and the transmembrane domain ligand structure generally refers to a molecular structure which can have recognition capacity on the transmembrane domain of the smooth receptor and can be combined with the transmembrane domain of the smooth receptor, and the recognition and the combination can be specific. The transmembrane domain ligand structure may be an inhibitor structure, an antagonist structure or an agonist structure. The transmembrane domain ligand structure can be an amide or imine structure containing a plurality of aliphatic rings, nitrogen-containing aliphatic rings, benzene rings and nitrogen-containing aromatic ring systems, for example, B can be selected from the following groups:

wherein J is selected from substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, substituted or unsubstituted pyridyl, and substituted or unsubstituted C3-C9 cycloalkyl, in the group J, the phenyl, benzyl, pyridyl and cycloalkyl can be monosubstituted or polysubstituted, and the substituents are respectively and independently selected from C1-C20, C1-C12, C1-C6, C1-C3 aliphatic group, halogen atoms such as F, Cl, Br, I and the like, nitro, amino, hydroxyl, alkoxy, cycloalkyl, C1-C20, C1-C12, C1-C6 aryl, C1-C20, C1-C12 and C1-C6 heteroaryl;

l is selected from hydrogen, C1-C20, C1-C12, C1-C6 and C1-C3 alkyl, in the group L, the alkyl can be monosubstituted or polysubstituted, and the substituent of the alkyl is selected from-NR 'R' substituted or unsubstituted C3-C9 cycloalkyl, wherein R 'and R' are respectively and independently selected from C1-C6 and C1-C3 alkyl;

m is selected from C1-C20, C1-C12, C1-C6 aryl or C1-C20, C1-C12, C1-C6 heteroaryl, in the group M, the aryl or heteroaryl can be monosubstituted or polysubstituted, and the substituents are selected from halogen atoms such as F, Cl, Br, I and the like, C1-C6 alkyl, C1-C6 alkoxy and-CF₃、-OCF₃And an alkylsulfonyl group. More specifically, there may be mentioned, but not limited to, the group structures corresponding to SAG (CAS number: 912545-86-9), LY2940680(CAS number: 1258861-20-9), GDC0449 (CAS number: 879085-55-9), LDE225(CAS number: 956697-53-3), SANT1(CAS number: 304909-07-7), SANT2(CAS number: 329196-48-7), and the like. In other embodiments of the present invention, B may be selected from the group shown below:

in the present invention, the aliphatic group generally refers to a chain hydrocarbon compound, and may be, for example, an alkyl group, an alkenyl group and an alkynyl group. The aliphatic group may be, but is not limited to, methyl, ethyl, ethenyl, ethynyl, propyl, propenyl, propynyl, butyl, butenyl, butynyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, and the like. The aliphatic group may contain a heteroatom, for example, the heteroatom may be N, S, P, O or the like.

In the present invention, the cycloalkyl group is understood to mean saturated and unsaturated (but not aromatic) cyclic hydrocarbons including, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. For cycloalkyl, also included are saturated cycloalkyl groups, wherein optionally at least one carbon atom may be replaced by a heteroatom, a preferred heteroatom may be S, N, P or O, and the like. The monounsaturated or polyunsaturated (preferably monounsaturated) cycloalkyl group having no heteroatom in the ring also belongs to the cycloalkyl group as long as it is not an aromatic system.

In the present invention, the alkoxy group includes, but is not limited to, methoxy, ethoxy, propoxy, and the like. The alkoxy group may be mono-substituted or poly-substituted, the substituent of the alkoxy group includes but is not limited to halogen atoms such as F, Cl, Br, etc., and the substituted alkoxy group may include but is not limited to fluoropropoxy, trifluoroethoxy, etc.

In the present disclosure, the aryl group generally refers to a closed ring system having at least one aromatic ring, generally excluding heteroatoms (e.g., N, O, etc. the aryl group includes, but is not limited to, phenyl, naphthyl, fluoranthenyl, fluorenyl, tetrahydronaphthyl, indanyl, anthracenyl, phenanthrenyl, dihydrophenanthrenyl, and the like.

In the present invention heteroaryl is understood to mean a heterocyclic ring system having at least one aromatic ring and optionally containing one or more heteroatoms selected from N, O, S and the like, said heteroaromatic ring groups including but not limited to furyl, thienyl, pyrrolyl, pyrazolyl, oxazolyl, thiazolyl, triazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, benzofuryl, benzothienyl, indolyl, isoindolyl, benzimidazolyl, indazolyl, benzothiazolyl, benzoxazolyl, quinolyl, isoquinolyl, quinoxalinyl, phthalazinyl, quinazolinyl, cinnolinyl, purinyl, carbazolyl, dibenzofuryl, acridinyl, phenazinyl, phenoxazine, phenothiazine, phenoxathiin and the like.

In the smooth receptor ligands provided herein, the linker may be generally linear or inactive to the smooth receptor. The length of the linker, which generally may be a polyethylene glycol oligomer (PEG oligomer with triazole) having a triazole structure, has an effect on the activity of the ligand, and in some embodiments of the present invention, the structural formula of the linker is as follows:

wherein m and n are each independently selected from integers between 0 and 7, for example, m can be 0, 1,2, 3,4, 5, 6 or 7, and for example, n can be 0, 1,2, 3,4, 5, 6 or 7.

In some embodiments of the invention, the smoothing receptor ligand may be specifically selected from compounds of the structural formula:

in another aspect, the present invention provides the use of the smoothing receptor ligand or its isomer, prodrug, solvate, or pharmaceutically acceptable salt thereof in the preparation of smooth receptor (SMO receptor) agonists, inhibitors or antagonists. An agonist generally refers to a class of substances that promote a molecular activity, a response, or a pathway, and an inhibitor generally refers to a class of substances that decrease or block a molecular activity, a response, or a pathway. The antagonist generally refers to a substance that blocks an agonist-mediated effect of another substance upon binding to a receptor and does not itself cause the biological effect to which the substance corresponds. When the length of the linker is within a certain range, the linker-optimized bipitch molecule acts on the receptor in a mode of simultaneously binding two binding domains of the smooth receptor, and achieves higher activity through a synergistic effect, wherein the achieved activity type is generally related to the activity type of the ligand, for example, the achieved activity effect can be the effects of agonism, inhibition, antagonism and the like.

In another aspect, the present invention provides a pharmaceutical composition comprising a smooth receptor ligand as described above, or an isomer, prodrug, solvate, or pharmaceutically acceptable salt thereof. In the pharmaceutical composition, the smooth receptor ligand or an isomer, a prodrug, a solvate or a pharmaceutically acceptable salt thereof can be used as the only active ingredient, and can also be combined with at least one or more other active ingredients for combined administration.

The pharmaceutical composition provided by the invention can also comprise pharmaceutically acceptable auxiliary materials or additives, and the auxiliary materials or additives can be selected from carriers, excipients, supporting materials, auxiliaries, lubricants, fillers, solvents, diluents, coloring agents, flavoring agents and the like.

The pharmaceutical composition according to the invention may be adapted for administration in any form, be it oral or parenteral, in particular pulmonary, nasal, rectal and/or intravenous injection, etc., and may for example be suitable for topical or systemic application, in particular intradermal, subcutaneous, intramuscular, intra-articular, intraperitoneal, pulmonary, buccal, sublingual, buccal, nasal, transdermal, vaginal, oral or parenteral application, etc.

The novel double-headed small molecule ligand for the smooth receptor provided by the invention is combined with the crystal structure data of the smooth receptor, and a connector is introduced into a proper site of an ectodomain ligand and a transmembrane domain ligand to obtain a brand-new double-headed ligand small molecule, so that the interaction between the ligand and the receptor and the biological activity of the ligand are enhanced. The small molecular ligand can be used as a tool molecule for the function and structure research of a smooth receptor, and can also be used as a medicine development candidate of diseases related to the smooth receptor.

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.

Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments, and is not intended to limit the scope of the present invention; in the description and claims of the present application, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.

Unless otherwise indicated, the experimental methods, detection methods, and preparation methods disclosed herein all employ techniques conventional in the art of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related arts. These techniques are well described in the literature, and may be found in particular in the study of the MOLECULAR CLONING, Sambrook et al: a LABORATORY MANUAL, Second edition, Cold Spring harbor LABORATORY Press, 1989 and Third edition, 2001; ausubel et al, Current PROTOCOLS Inmolecular BIOLOGY, John Wiley & Sons, New York, 1987 and periodic updates; the series METHODS IN ENZYMOLOGY, Academic Press, San Diego; wolffe, CHROMATINSTRUCUTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998; (iii) Methods Inenzymolygy, Vol.304, Chromatin (P.M. Wassarman and A.P.Wolffe, eds.), academic Press, San Diego, 1999; and METHODS IN MOLECULAR BIOLOGY, Vol.119, chromatography protocols (P.B.Becker, ed.) Humana Press, Totowa, 1999, etc.