CN114262319A

CN114262319A - Bifunctional molecules, preparation method and application thereof

Info

Publication number: CN114262319A
Application number: CN202011386126.9A
Authority: CN
Inventors: 杨光; 李文龙
Original assignee: Nanchang Aorui Pharmaceutical Co ltd
Current assignee: Nanjing Aorui Pharmaceutical Co ltd
Priority date: 2020-12-01
Filing date: 2020-12-01
Publication date: 2022-04-01
Anticipated expiration: 2040-12-01
Also published as: CN114262319B

Abstract

The invention discloses a bifunctional molecule, a preparation method and application thereof. The bifunctional molecule of the invention can inhibit the proliferation of LNCap cells, and part of the compounds also have the effect of inhibiting the proliferation of PC-3 cells, so that the bifunctional molecule in the application has the potential of treating cancers, particularly prostate cancer. In addition, the preparation method of the bifunctional molecule has the advantages of mild conditions, simplicity in operation, no high-temperature and low-temperature extreme reaction conditions, no use of high-risk highly toxic substances, suitability for laboratory synthesis and industrial production, and wide industrial application prospect.

Description

Bifunctional molecules, preparation method and application thereof

Technical Field

The invention relates to the field of pharmaceutical combination, in particular to bifunctional molecules, a preparation method and application thereof.

Background

Prostate cancer is a common male malignancy, with a worldwide incidence second only to lung cancer, ranking second to all male malignancies. The incidence of Chinese prostate cancer is gradually increased year by year, and the prostate cancer is predicted to become the third disease of cancer death of men in China by 2020. Androgen Deprivation Therapy (ADT) is currently the primary treatment for early stage prostate cancer, including surgical and drug castration. But ADT does not cure prostate cancer, and after a median treatment period of 14-30 months, almost all patients gradually develop castration-resistant prostate cancer (CRPC), patients are no longer sensitive to ADT, and median survival is less than 20 months. In prostate cancer tissue, proliferation or mutation of the Androgen Receptor (AR) can sensitize it to lower levels of androgens in serum, a major cause of prostate cancer progression. At present, the androgen receptor inhibitor for treating CRPC represents the drugs such as abiraterone and enzalutamide. However, 15-25% of patients do not respond to second generation hormone therapy such as abiraterone and enzalutamide, and most of the patients with reactivity eventually develop severe drug resistance, resulting in poor prognosis.

Ubiquitin-proteasome system (UPS) is the major pathway for protein degradation in cells, involving more than 80% of the proteins in cells. UPS is composed of ubiquitin (Ub), ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2), ubiquitin-protein ligase (E3), 26S proteasome and its target protein. The ubiquitin activating enzyme system is responsible for activating ubiquitin and binding it to the protein to be degraded, forming the target protein polyubiquitin chain, i.e. ubiquitination. The proteasome system can recognize ubiquitinated proteins and degrade them. Proteolytic targeting chimeric molecules (PROTACs) are hybrid bifunctional small molecule compounds comprising a small molecule compound capable of binding to a target protein of interest, a linker at a suitable position, and a small molecule compound capable of binding to E3 ubiquitin ligase. The PROTACs form a ternary polymer of a target protein-PROTACs-E3 by drawing a target protein of interest and intracellular E3, apply a ubiquitinated protein tag to the target protein of interest through E3 ubiquitin ligase, and specifically degrade the target protein by using a ubiquitin-proteasome system (UPS).

In 2018, Arvinus discloses a class of androgen receptor targeting degraders (US 2018/0099940a1), which represent the drug ARV-110, currently in phase I clinical for the treatment of CRPC. Clinical phase I data results also indicate that ARV-110 is safe and effective in patients with metastatic castration resistant prostate cancer (mCRPC).

Disclosure of Invention

The invention aims to solve the technical problem that the conventional androgen receptor inhibitor has a single structure, and therefore, the invention provides bifunctional molecules completely different from the prior art, a preparation method and application thereof. The bifunctional molecule can inhibit the proliferation of LNCap cells, and part of the compound also has the effect of inhibiting the proliferation of PC-3 cells.

The invention provides a compound shown as a formula I or a pharmaceutically acceptable salt thereof;

wherein:

is C₄-C₇Cycloalkylene radicals or "substituted by one or more C₁-C₆Alkyl substituted C₄-C₇Cycloalkylene radicals;

is phenylene or "5-6 membered heteroarylene having 1,2 or 3 heteroatoms independently selected from one or more of N, O and S", the a terminus is attached to the carbonyl group and the b terminus is attached to X;

is phenylene, substituted by one or more R¹A substituted phenylene, "a 5-6 membered heteroarylene having 1,2, or 3 heteroatoms independently selected from one or more of N, O and S," or "substituted with one or more R²A 5-6 membered heteroarylene group having 1,2 or 3 substituted atoms, a heteroatom independently selected from one or more of N, O and S, the c-terminus is attached to X, and the d-terminus is attached to Y;

R¹is halogen, C₁-C₆Alkyl or C₁-C₆An alkoxy group;

R²is halogen, C₁-C₆Alkyl or C₁-C₆An alkoxy group;

x is O, S, CH₂Or NR³；R³Is hydrogen or C₁-C₆An alkyl group;

y is CH or N, R' is hydrogen or C₁-C₆An alkyl group;

n is 0 or 1, and when n is 0, Y is absent;

z is O or S;

r is hydrogen or C₁-C₆An alkyl group;

or, R and R' are linked to form- (CH)₂)_m-structure, m is 2, 3 or 4.

In a preferred embodiment, in the compound represented by formula I or the pharmaceutically acceptable salt thereof, some groups may be defined as follows, and the rest groups may be defined as described in any one of the above (for this expression, hereinafter, simply referred to as "in a preferred embodiment"):

when in use

Is formed by one or more C₁-C₆Alkyl substituted C₄-C₇When cycloalkylene, said C₁-C₆The alkyl group is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group or a tert-butyl group, and more preferably a methyl group; said C₄-C₇The cycloalkylene group is preferably a cyclobutyl group, a cyclopentyl group, a cyclohexylene group or a cycloheptylene group, and more preferably

Most preferably

The plurality is two, three or four.

In a preferred embodiment, when

To be covered with a plurality of C₁-C₆Alkyl substituted C₄-C₇When cycloalkylene, said group is substituted by more than one C₁-C₆Alkyl substituted C₄-C₇Cycloalkylene is

In a preferred embodiment, when

Is C₄-C₇When cycloalkylene, said C₄-C₇The cycloalkylene group is preferably a cyclobutyl group, a cyclopentyl group, a cyclohexylene group or a cycloheptylene group, and more preferably

Most preferably

In a preferred embodiment of the present invention,

preferably by a plurality of C₁-C₆Alkyl substituted C₄-C₇Cycloalkylene or C₄-C₇A cycloalkylene group; more preferably

In a preferred embodiment, when

When it is a phenylene group, the phenylene group is preferably

More preferably

In a preferred embodiment, when

In the case of "a 5-6 membered heteroarylene group having 1,2 or 3 hetero atoms independently selected from one or more of N, O and S", the 5-6 membered heteroarylene group is preferably a triazolylene group, a pyrazolyl arylene group, a pyridyl group, a pyrimidylene group, a pyrazinylene group, an oxazolylene group or a pyridazinylene group, more preferably a triazolylene group, a pyrazolyl group, a pyridyl group, a pyrimidylene group, a pyrazinylene group, an oxazolylene group or a pyridazinylene group

In a preferred embodiment of the present invention,

preference is given to

In a preferred embodiment, when

When it is a phenylene group, the phenylene group is preferably

More excellentSelecting

In a preferred embodiment, when R¹In the case of halogen, the halogen is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine, chlorine or bromine.

In a preferred embodiment, when R¹Is C₁-C₆When alkyl, said C₁-C₆The alkyl group is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group or a tert-butyl group, and more preferably a methyl group.

In a preferred embodiment, when R¹Is C₁-C₆At alkoxy, said C₁-C₆The alkoxy group is preferably a methoxy group, an ethoxy group, an n-propoxy group, an isopropoxy group, an n-butoxy group, an isobutoxy group, a sec-butoxy group or a tert-butoxy group, and more preferably a methoxy group.

In a preferred embodiment, when

Is represented by one or more R¹In the case of substituted phenylene, the phenylene group is preferably

More preferably

Said plurality is preferably two or three.

In a preferred embodiment, when

Is to be an R¹In the case of substituted phenylene radicals, said radical is substituted by one R¹Substituted phenylene radicals are preferred

More preferably

In a preferred embodiment, when

In the case of "a 5-6 membered heteroarylene group having 1,2 or 3 hetero atoms independently selected from one or more of N, O and S", the 5-6 membered heteroarylene group is preferably a triazolylene group, a pyrazolyl arylene group, a pyridyl ene group, a pyrimidylene group, a pyrazinylene group, an oxazolylene group and a pyridazinylene group, more preferably a triazolylene group, a pyrazolyl ene group, a pyridyl ene group, a pyrimidylene group, a pyrazinylene group, an oxazolylene group and a pyridazinylene group

In a preferred embodiment, when R²In the case of halogen, the halogen is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine, chlorine or bromine.

In a preferred embodiment, when R²Is C₁-C₆When alkyl, said C₁-C₆The alkyl group is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group or a tert-butyl group, and more preferably a methyl group.

In a preferred embodiment, when R²Is C₁-C₆At alkoxy, said C₁-C₆The alkoxy group is preferably a methoxy group, an ethoxy group, an n-propoxy group, an isopropoxy group, an n-butoxy group, an isobutoxy group, a sec-butoxy group or a tert-butoxy group, and more preferably a methoxy group.

In a preferred embodiment, when

Is "by one or more R²When the substituted 5-6 membered heteroarylene group "has 1,2 or 3 substituted atoms and one or more heteroatoms independently selected from N, O and S, the 5-6 membered heteroarylene group is preferably a triazylene groupOxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinylene, oxazolylene or pyridazinylene, and

in a preferred embodiment of the present invention,

preferably phenylene, by an R¹Substituted phenylene or "5-6 membered heteroarylene having 1,2 or 3 heteroatoms independently selected from one or more of N, O and S", more preferably

In a preferred embodiment, when R³Is C₁-C₆When alkyl, said C₁-C₆The alkyl group is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group or a tert-butyl group, and more preferably a methyl group.

In a preferred embodiment, when R' is C₁-C₆When alkyl, said C₁-C₆The alkyl group is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group or a tert-butyl group, and more preferably a methyl group.

In a preferred embodiment, when R is C₁-C₆When alkyl, said C₁-C₆The alkyl group is preferably a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group or a tert-butyl group, and more preferably a methyl group.

In a preferred embodiment of the present invention,

is formed by one or more C₁-C₆Alkyl substituted C₄-C₇Cycloalkylene, preferably

In a preferred embodiment of the present invention,

is phenylene, preferably

In a preferred embodiment of the present invention,

is phenylene, "substituted by one or more R¹Substituted phenylene "or" 5-6 membered heteroarylene having 1,2 or 3 heteroatoms independently selected from one or more of N, O and S ", preferably phenylene or" substituted with one or more R "groups¹Substituted phenylene ".

In a preferred embodiment, R¹Is hydrogen.

In a preferred embodiment, R' is hydrogen.

In a preferred embodiment, n is 1.

In a preferred embodiment, R is hydrogen.

In a preferred embodiment of the present invention,

is composed of

In a preferred embodiment, the compound of formula I is any of the following:

the first scheme is as follows:

to be covered with a plurality of C₁-C₆Alkyl substituted C₄-C₇Cycloalkylene or C₄-C₇Cycloalkylene, preferably

Is phenylene, 5-6 membered heteroarylene with 1,2 or 3 heteroatoms independently selected from N, O and S, wherein the 5-6 membered heteroarylene is pyridazinylene, preferably

Is phenylene, "substituted by one or more R¹Substituted phenylene "or" 5-6 membered heteroarylene having 1,2 or 3 heteroatoms independently selected from one or more of N, O and S ", said phenylene preferably being

The 5-to 6-membered heteroarylene group is preferred

Scheme II:

Is phenylene or "substituted by one or more R¹Substituted phenylene ", said phenylene preferably being

The third scheme is as follows:

Is phenylene, preferably

R¹Is halogen or C₁-C₆An alkyl group;

n is 1;

y is N and R' is hydrogen.

In a preferred embodiment, the compound represented by formula I is any one of the following compounds:

the invention also provides a preparation method of the compound shown in the formula I, which is any one of the following schemes:

in scheme A, when n is 0 or n is 1 and Y is CH, the preparation method of the compound shown in the formula I is as follows:

in a solvent, in the presence of alkali and a dehydrating agent, carrying out dehydration reaction as shown in the following on a compound shown in a formula II and a compound shown in a formula III to obtain a compound shown in a formula I;

scheme B, when N is 1 and Y is N, the preparation of the compound of formula I is as follows:

in a solvent, in the presence of a condensing agent, carrying out a condensation reaction shown in the following formula on a compound shown in a formula IV and a compound shown in a formula III to obtain a compound shown in a formula I;

in the scheme a, the conditions and steps of the dehydration reaction are those conventional in the art, and are preferably as follows:

in the dehydration reaction, the solvent may be a solvent conventional in the art for such dehydration reactions, preferably an amide-based solvent, more preferably N, N-dimethylformamide.

In the dehydration reaction, the dehydrating agent may be a dehydrating agent conventional in the dehydration reaction of this type in the art, preferably HATU.

In the dehydration reaction, the base may be a base conventional in the art for such dehydration reactions, preferably N, N-diisopropylethylamine or triethylamine.

In said scheme B, the conditions and steps of the condensation reaction are those conventional in the art, and are preferably as follows:

in the condensation reaction, the solvent may be a solvent conventional in the art for such condensation reactions, preferably an ethereal solvent, more preferably tetrahydrofuran.

In the condensation reaction, the condensing agent may be a condensing agent conventional in the art for such condensation reactions, and is preferably thiocarbonyldiimidazole or N, N' -carbonyldiimidazole.

In the condensation reaction, the reaction temperature may be a temperature conventional in the art for such condensation reactions, for example, 10 ℃ to 30 ℃.

Scheme a of the above-described process for the preparation of the compound of formula I may further comprise the steps of: in a solvent, in the presence of a deprotection reagent, carrying out deprotection reaction on a compound shown as a formula V to obtain a compound shown as a formula II;

R⁴is C₁-C₃An alkyl group.

The conditions and procedures for the deprotection reaction are those conventional in the art, and are preferably as follows:

in the deprotection reaction, the solvent may be a solvent conventional in the art for such deprotection reactions, preferably an alcohol solvent and/or water, more preferably methanol and/or water.

In the deprotection reaction, the solvent of the deprotection reagent can be a deprotection reagent which is conventional in the deprotection reactions of the type in the field, and sodium hydroxide is preferred.

In the deprotection reaction, R⁴Preferably methyl, ethyl, propyl or isopropyl, more preferably methyl.

Scheme a of the above-described process for the preparation of the compound of formula I may further comprise the steps of: in a solvent, in the presence of acid, carrying out substitution reaction shown as the following on a compound shown as a formula VII and a compound shown as a formula VI to obtain a compound shown as a formula V;

R⁵is halogen.

The conditions and procedures of the substitution reaction are those conventional in the art, and are preferably as follows:

in the substitution reaction, the solvent can be a solvent conventional in the substitution reaction of the type in the field, preferably an ether solvent, and more preferably dioxane.

In the substitution reaction, the acid may be an acid conventional in the art for such substitution reactions, preferably hydrochloric acid.

In the substitution reaction, the R is⁵Preferably fluorine, chlorine, bromine or iodine, more preferably chlorine or bromine.

Scheme a of the above-described process for the preparation of the compound of formula I may further comprise the steps of: in a solvent, in the presence of alkali and a condensing agent, carrying out acylation reaction as shown in the specification on a compound shown in a formula VIII and a compound shown in a formula VIIII to obtain a compound shown in a formula VII;

the conditions and procedures of the acylation reaction are those conventional in the art, and are preferably as follows:

in the acylation reaction, the solvent may be a solvent conventional in the art for such acylation reactions, preferably an amide solvent, more preferably N, N-dimethylformamide.

In the acylation reaction, the condensing agent can be a condensing agent conventional in the acylation reaction of the type in the art, and HATU is preferred.

In the acylation reaction, the base can be a base conventional in the art for such acylation reactions, preferably N, N-diisopropylethylamine or triethylamine.

Scheme B of the above process for the preparation of the compound of formula I may further comprise the steps of: in a solvent, in the presence of hydrogen and a catalyst, carrying out a reduction reaction shown as the following on the compound A to obtain a compound shown as a formula IV;

the conditions and procedures of the reduction reaction are conventional in the art and are preferably as follows:

in the reduction reaction, the solvent may be a solvent conventional in the art for such reduction reactions, preferably an alcohol solvent and/or an ether solvent, more preferably tetrahydrofuran and/or methanol.

In the reduction reaction, the catalyst may be a catalyst conventional in the art for such reduction reactions, preferably palladium on carbon and/or iron powder.

Scheme B of the above process for the preparation of the compound of formula I may further comprise the steps of: when X is NH, carrying out nucleophilic substitution reaction shown in the specification on the compound B and the compound C in a solvent in the presence of acid to obtain a compound A;

R⁶is halogen.

The conditions and procedures for the nucleophilic substitution reaction are those conventional in the art, and are preferably as follows:

in the nucleophilic substitution reaction, the solvent may be a solvent conventional in such nucleophilic substitution reactions in the art, preferably an ether solvent, more preferably dioxane.

In the nucleophilic substitution reaction, the R⁶Preferably fluorine, chlorine, bromine or iodine, more preferably chlorine or bromine.

In the nucleophilic substitution reaction, the acid may be an acid conventional in such nucleophilic substitution reactions in the art, preferably hydrochloric acid.

Scheme B of the above process for the preparation of the compound of formula I may further comprise the steps of: in a solvent, in the presence of hydrogen and a catalyst, carrying out a hydrogenation reaction on the compound D as shown in the specification to obtain a compound B;

the conditions and procedures of the hydrogenation reaction are those conventional in the art, and are preferably as follows:

in the hydrogenation reaction, the solvent may be a solvent conventional in the art for such hydrogenation reactions, preferably an alcohol solvent and/or an ether solvent, more preferably tetrahydrofuran and/or methanol.

In the hydrogenation, the catalyst may be a catalyst conventional in the art for such hydrogenation, preferably palladium on carbon and/or iron powder.

Scheme B of the above process for the preparation of the compound of formula I may further comprise the steps of: in a solvent, in the presence of alkali and a condensing agent, carrying out dehydration condensation reaction on the compound E and the compound F as shown in the specification to obtain a compound D;

the conditions and steps of the dehydration condensation reaction are those conventional in the art, and are preferably as follows:

in the dehydration condensation reaction, the solvent may be a solvent conventional in the dehydration condensation reaction of this type in the art, preferably an amide solvent, more preferably N, N-dimethylformamide.

In the dehydration condensation reaction, the condensing agent may be a condensing agent conventional in the art for such dehydration condensation reaction, preferably HATU.

In the dehydration condensation reaction, the base may be a base conventional in the dehydration condensation reaction of this type in the art, and preferably N, N-diisopropylethylamine or triethylamine.

Scheme B of the above process for the preparation of the compound of formula I may further comprise the steps of: in a solvent, in the presence of alkali, carrying out an ether forming reaction shown as the following on a compound F and a compound G to obtain a compound E;

R⁷is halogen.

The conditions and procedures of the ether-forming reaction are those conventional in the art, and are preferably as follows:

in the ether-forming reaction, the solvent may be a solvent conventional in the art for such ether-forming reaction, preferably an ethereal solvent, more preferably tetrahydrofuran.

In the ether-forming reaction, the R⁷Preferably fluorine, chlorine, bromine or iodine, more preferably chlorine or bromine.

In the ether-forming reaction, the base may be a base conventional in the art for such ether-forming reactions, preferably sodium hydrogen.

The invention also provides a pharmaceutical composition, which comprises a substance X and a pharmaceutic adjuvant, wherein the substance X is the compound shown in the formula I or the pharmaceutically acceptable salt thereof. The substance X is preferably used in a therapeutically effective amount.

The invention also provides application of the substance X or the pharmaceutical composition in preparing LNCap cell and/or PC-3 cell proliferation inhibitor.

The invention also provides the use of substance X as defined above or a pharmaceutical composition as defined above for the manufacture of a medicament for the treatment and/or prevention of a condition associated with the androgen receptor, such as a cancer, for example prostate cancer.

Unless otherwise indicated, the following terms appearing in the specification and claims of the invention have the following meanings:

the medicinal auxiliary materials can be auxiliary materials widely adopted in the field of medicine production. The excipients are used primarily to provide a safe, stable and functional pharmaceutical composition and may also provide methods for dissolving the active ingredient at a desired rate or for promoting the effective absorption of the active ingredient after administration of the composition by a subject. The pharmaceutical excipients may be inert fillers or provide a function such as stabilizing the overall pH of the composition or preventing degradation of the active ingredients of the composition. The pharmaceutical excipients may include one or more of the following excipients: buffers, chelating agents, preservatives, co-solvents, stabilizers, excipients and surfactant colorants, flavors and sweeteners.

The term "pharmaceutically acceptable salt" refers to salts prepared from the compounds of the present invention with relatively nontoxic, pharmaceutically acceptable acids or bases. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral forms of such compounds with a sufficient amount of a pharmaceutically acceptable base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to: lithium salt, sodium salt, potassium salt, calcium salt, aluminum salt, magnesium salt, zinc salt, bismuth salt, ammonium salt, and diethanolamine salt. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable acid in neat solution or in a suitable inert solvent. The pharmaceutically acceptable acids include inorganic acids including, but not limited to: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, sulfuric acid, and the like. The pharmaceutically acceptable acids include organic acids including, but not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acid citric acid, oleic acid, tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentisic acid, fumaric acid, gluconic acid, saccharic acid, formic acid, ethanesulfonic acid, pamoic acid (i.e. 4, 4' -methylene-bis (3-hydroxy-2-naphthoic acid)), amino acids (e.g. glutamic acid, arginine), and the like. When the compounds of the present invention contain relatively acidic and relatively basic functional groups, they may be converted to base addition salts or acid addition salts. See in particular Berge et al, "Pharmaceutical Salts", Journal of Pharmaceutical Science 66:1-19(1977), or, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P.Heinrich Stahl and Camile G.Wermuth, ed., Wiley-VCH, 2002).

When any variable (e.g. R)¹) In the definition of a compound, the occurrence at each position of the variable is defined multiple times independently of the occurrence at the remaining positions, and their meanings are independent of each other and independent of each other. Thus, if a group is substituted by 1,2 or 3R¹Substituted by radicals, i.e. the radical may be substituted by up to 3R¹Substituted in the position R¹Is defined by the definition of (1) and the remaining positions R¹Are defined independently of each other. In addition, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

The term "alkyl" refers to a straight or branched chain alkyl group having the indicated number of carbon atoms.

The term "alkoxy" refers to the group-O-R^XWherein R is^XIs an alkyl group as defined above.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "heteroaryl" refers to aryl groups containing 1,2, 3, or 4 heteroatoms independently selected from N, O and S, which are monocyclic aromatic systems, such as furyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, and the like.

The term "treatment" or its equivalent when used with reference to, for example, cancer, refers to a procedure or process for reducing or eliminating the number of cancer cells in a patient or alleviating the symptoms of cancer. "treating" cancer or another proliferative disorder does not necessarily mean that the cancer cells or other disorder will actually be eliminated, that the number of cells or disorders will actually be reduced or that the symptoms of the cancer or other disorder will actually be alleviated. Generally, methods for treating cancer are performed even with a low likelihood of success, but are still considered to induce an overall beneficial course of action, given the patient's medical history and estimated survival expectations.

The term "prevention" refers to a reduced risk of acquiring or developing a disease or disorder.

In the above-mentioned applications, the cell proliferation inhibitor can be used in a mammalian organism; also useful in vitro, primarily for experimental purposes, for example: the kit can be used as a standard sample or a control sample for comparison, or can be prepared into a kit according to the conventional method in the field, so as to provide rapid detection for the cell inhibition effect.

The above preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention without departing from the common general knowledge in the art.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows: the bifunctional molecule can inhibit the proliferation of LNCap cells, and part of the compound has the effect of inhibiting the proliferation of PC-3 cells.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.

Example 1: synthesis of intermediate 1

Tert-butyl ((1r,3r) -3-hydroxy-2, 2,4, 4-tetramethylcyclobutane) carbamate (3.1g, 12.86mmol, 1eq) was dissolved in anhydrous THF, sodium hydride (617mg, 15.43mmol, 60%, 1.2eq) was added at 0 deg.C, and after stirring under nitrogen for 30min, 2-chloro-5-fluorobenzonitrile (2g, 12.86mmol, 1eq) was added and stirred at room temperature for 4 h. After the TLC detection reaction, water was added to quench the reaction, THF was removed under reduced pressure, the reaction mixture was diluted with ethyl acetate (100mL), washed with water (50mL), washed with saturated brine (50mL), dried over anhydrous sodium sulfate and concentrated, and the residue was subjected to column chromatography (PE/EA 10:1) to give 3.5g of a product. The above product was dissolved in 50mL dioxane, HCl dioxane solution (4M,10mL) was added dropwise, stirred overnight at room temperature, filtered, filter cake washed with dioxane, ether, and dried to give 3.6g white solid. ESI MS M/z 280[ M + H ]]⁺. The above product (200mg, 0.72mmol, 1eq), p-nitrobenzoic acid (144mg, 0.86mmol, 1.2eq) were dissolved in DMF (15mL), DIEA (179. mu.L, 1.08mmol, 1.5eq) and HATU (410mg, 1.08mmol, 1.5eq) were added, respectively, and stirred at room temperature for 2 h. After the TLC detection reaction, water (50mL) was added for dilution, ethyl acetate (50mL) was extracted three times, the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography to give 280mg of a product. Dissolving the above product in THF, adding palladium carbon, stirring at room temperature under hydrogen condition overnight, vacuum filtering, concentrating the filtrate to obtain product 260mg, ESI MS M/z 399[ M + H ]]⁺. The above product (100mg, 0.26mmol, 1eq) was dissolved in DMSO, 3-chloro-4-nitrobenzene (54mg, 0.3mmol, 1.2eq) and DIEA (64. mu.L, 0.38mmol, 1.5eq) were added, respectively, and stirred at 120 ℃ overnight. After completion of the TLC detection reaction, the mixture was diluted with ethyl acetate (20mL), washed with water (20mL), and saturated brine (20mL)Washed, dried over anhydrous sodium sulfate, concentrated and the residue subjected to column chromatography to give 120mg of a product. Dissolving the above products in THF, adding palladium carbon, stirring at room temperature under hydrogen condition overnight, vacuum filtering, concentrating the filtrate to obtain 124mg of product ESI MS M/z 523[ M + H ]]⁺。

Example 2: synthesis of intermediate 2

The above product (compound 1-4) (50mg, 0.13mmol, 1eq) was dissolved in DMSO, 4-fluoronitrobenzene (21mg, 0.15mmol, 1.2eq) and DIEA (31. mu.L, 0.19mmol, 1.5eq) were added, respectively, and stirred at 120 ℃ overnight. After the TLC detection reaction, the reaction mixture was diluted with ethyl acetate (20mL), washed with water (20mL), washed with saturated brine (20mL), dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography to give 28mg of the product. Dissolving the above product in THF, adding palladium carbon, stirring at room temperature under hydrogen condition overnight, vacuum filtering, concentrating the filtrate to obtain 25mg of product ESI MS M/z 490[ M + H ]]⁺。

Example 3: synthesis of intermediate 3

The above product (compound 1-4) (50mg, 0.13mmol, 1eq) was dissolved in DMSO, 3, 4-difluoronitrobenzene (24mg, 0.15mmol, 1.2eq) and DIEA (31. mu.L, 0.19mmol, 1.5eq) were added, respectively, and 120 ℃ was stirred overnight. After the TLC detection reaction, ethyl acetate (20mL) was added for dilution, washed with water (20mL), washed with saturated brine (20mL), dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography to give 36mg of the product. Dissolving the above products in THF, adding palladium carbon, stirring at room temperature under hydrogen condition overnight, vacuum filtering, concentrating the filtrate to obtain 35mg of product ESI MS M/z 508[ M + H ]]⁺。

Example 4: synthesis of intermediate 4

The above product (compound 1-4) (50mg, 0.13mmol, 1eq) was dissolved in DMSO, 3, 4-difluoronitrobenzene (23mg, 0.15mmol, 1.2eq) and DIEA (31. mu.L, 0.19mmol, 1.5eq) were added, respectively, and 120 ℃ was stirred overnight. After the TLC detection reaction, ethyl acetate (20mL) was added for dilution, washed with water (20mL), washed with saturated brine (20mL), dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography to give 45mg of the product. Dissolving the above product in THF, adding palladium carbon, stirring at room temperature under hydrogen condition overnight, vacuum filtering, concentrating the filtrate to obtain product 40mg, ESI MS M/z 504[ M + H ] (ESI MS M/z)]⁺。

Example 5: synthesis of intermediate 5

The above product (compound 1-4) (50mg, 0.13mmol, 1eq) was dissolved in DMSO, 3-bromo-4-fluoronitrobenzene (33mg, 0.15mmol, 1.2eq) and DIEA (31. mu.L, 0.19mmol, 1.5eq) were added, respectively, and stirred at 120 ℃ overnight. After completion of the TLC detection reaction, the reaction mixture was diluted with ethyl acetate (20mL), washed with water (20mL), washed with saturated brine (20mL), dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography to give 43mg of the product. Dissolving the above product in THF, adding palladium carbon, stirring at room temperature under hydrogen condition overnight, vacuum filtering, concentrating the filtrate to obtain 41mg of product ESI MS M/z 587[ M + H ]]⁺。

Example 6: synthesis of intermediate 6

Dissolving tert-butyl ((1r,4r) -4-hydroxycyclohexane carbamate (2.7g, 12.86mmol, 1eq) in anhydrous THF, adding sodium hydride (617mg, 15.43mmol, 60%, 1.2eq) at 0 ℃, stirring under nitrogen for 30min, adding 2-chloro-5-fluorobenzonitrile (2g, 12.86mmol, 1eq), stirring at room temperature for 4h, after TLC detection reaction, adding water to quench the reaction, removing THF under reduced pressure, adding ethyl acetate (100 g, 12.86mmol, 1eq), addingmL), washed with water (50mL), washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography (PE/EA 10:1) to give 3.0g of the product. The above product was dissolved in 50mL dioxane, HCl dioxane solution (4M,10mL) was added dropwise, stirred overnight at room temperature, filtered, filter cake washed with dioxane, ether, dried to give 2.6g white solid. ESI MS M/z 251[ M + H ]]⁺. The above product (200mg, 0.72mmol, 1eq), 6-aminopyridazine-3-carboxylic acid (133mg, 0.86mmol, 1.2eq) were dissolved in DMF (15mL), DIEA (179. mu.L mg, 1.08mmol, 1.5eq) and HATU (410mg, 1.08mmol, 1.5eq) were added, respectively, and stirred at room temperature for 2 h. After the TLC detection reaction, water (50mL) was added for dilution, ethyl acetate (50mL) was extracted three times, the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography to give 225mg of the product. Dissolving the above product (410mg, 1.08mmol, 1.5eq) in THF, adding palladium on carbon, stirring overnight at room temperature under hydrogen, vacuum filtering, concentrating the filtrate to obtain 180mg product, ESI MS M/z 372[ M + H ] 372]⁺. The above product (180mg, 0.48mmol, 1eq) was dissolved in anhydrous THF, sodium hydride (23mg, 0.58mmol, 60%, 1.2eq) was added at 0 deg.C, and after stirring under nitrogen for 30min, 3-chloro-4-nitrobenzene (84mg, 0.58mmol, 1eq) was added and stirred at room temperature for 4 h. After the TLC detection reaction, water was added to quench the reaction, THF was removed under reduced pressure, and the reaction mixture was diluted with ethyl acetate (20mL), washed with water (20mL), washed with saturated brine (20mL), dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography to give 75mg of the product. The above product (75mg, 0.14mmol, 1eq) was dissolved in a mixed solution (10mL) of methanol and saturated aqueous ammonium chloride (1: 1), iron powder was added, and stirring was carried out at 65 ℃ for 1 h. After completion of the TLC detection reaction, the reaction mixture was diluted with ethyl acetate (20mL), washed with water (20mL), washed with saturated brine (20mL), dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to column chromatography to give 652 mg of an intermediate. ESI MS M/z 498[ M + H ]]⁺。

Example 7: synthesis of intermediate 7

Mixing 4- ((1r,3r) -3-amino-2, 2,4, 4-tetraMethylcyclobutylalkoxy) -2-chlorobenzonitrile (100mg, 0.36mmol, 1eq) was dissolved in anhydrous DMF, pyrazole-3-carboxylic acid (40mg, 0.43mmol, 1.2eq) was dissolved in DMF (15mL), DIEA (89. mu.L, 0.54mmol, 1.5eq) and HATU (205mg, 0.54mmol, 1.5eq) were added, respectively, and stirred at room temperature for 2 h. After the TLC detection reaction, water (50mL) was added for dilution, ethyl acetate (50mL) was extracted three times, the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography to give 82mg of the product. The above product (82mg, 0.22mmol, 1eq) was dissolved in DMF (15mL), potassium carbonate (57mg, 0.33mmol, 1.5eq) and p-nitrobenzyl bromide (72mg, 0.33mmol, 1.5eq) were added, respectively, stirred at 120 ℃ for 2h, diluted with water (50mL), extracted with ethyl acetate (50mL) three times, the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography gave 55 mg. Dissolving the above product in THF, adding palladium carbon, stirring at room temperature under hydrogen condition overnight, vacuum filtering, concentrating the filtrate to obtain intermediate 745 mg, ESI MS M/z 479[ M + H ]]⁺。

Example 8: synthesis of intermediate 8

4- ((1r,3r) -3-amino-2, 2,4, 4-tetramethylcyclobutylalkoxy) -2-chlorobenzonitrile (100mg, 0.36mmol, 1eq), 1H-1,2, 4-triazole-3-carboxylic acid (40mg, 0.43mmol, 1.2eq) were dissolved in DMF (15mL), DIEA (89. mu.L, 0.54mmol, 1.5eq) and HATU (205mg, 0.54mmol, 1.5eq) were added, respectively, and the mixture was stirred at room temperature for 2 hours. After the TLC detection reaction, water (50mL) was added for dilution, ethyl acetate (50mL) was extracted three times, the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography to give 65mg of a product. The above product (65mg, 0.17mmol, 1eq) was dissolved in DMF (15mL), potassium carbonate (45mg, 0.26mmol, 1.5eq) and p-nitrobenzyl bromide (46mg, 0.26mmol, 1.5eq) were added respectively, stirred at 120 ℃ for 2h, diluted with water (50mL), extracted with ethyl acetate (50mL) three times, the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography gave 32 mg. Dissolving the above product in THF, adding palladium carbon,stirring overnight at room temperature under hydrogen, filtering, concentrating the filtrate to obtain intermediate 845 mg, ESI MS M/z 479[ M + H ]]⁺。

Example 9: synthesis of intermediate 9

Methyl 4-bromomethylbenzoate (500mg, 2.18mmol, 1eq) was dissolved in DMF (15mL), 4-Boc aminopiperidine (481mg, 2.40mmol, 1.1eq) and potassium carbonate (362mg, 2.62mmol, 1.2eq) were added, respectively, stirred at 120 ℃ for 2h, diluted with water (50mL), extracted with ethyl acetate (50mL) three times, the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography to give 620mg of the product. The above product (620mg, 1.78mmol, 1eq) was dissolved in a mixed solvent of methanol and 10% aqueous NaOH (1:1,20mL), reacted at room temperature under stirring for 2h, diluted with water (50mL), washed with ethyl acetate (50mL), the pH of the aqueous layer was adjusted to 4 with dilute hydrochloric acid, extracted with ethyl acetate (50mL), the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography to give 430mg of the product. The above product (266mg, 0.8mmol, 1eq), 4- (((1r,4r) -4-aminocyclohexylalkoxy) -2-chlorobenzonitrile (200mg, 0.8mmol, 1eq) was dissolved in DMF (15mL), DIEA (158. mu.L, 0.96mmol, 1.2eq) and HATU (365mg, 0.96mmol, 1.2eq) were added, respectively, stirred at room temperature for 2h after TLC detection, diluting with water (50mL), extracting with ethyl acetate (50mL) for three times, concentrating the organic layer, washing with saturated saline, drying over anhydrous sodium sulfate, concentrating under reduced pressure, performing column chromatography to obtain 185mg, dissolving the above product (185mg, 0.33mmol, 1eq) in dioxane (15mL), HCl in dioxane (4M,10mL) was slowly added dropwise at 0 deg.C, stirred overnight at room temperature, after TLC detection, concentrate under reduced pressure to give intermediate 9230 mg. ESI MS M/z 467[ M + H.]⁺。

Example 10: synthesis of intermediate 10

Dissolving the intermediate 1(35mg, 0.067mmol, 1eq) in THF (10mL), adding paraformaldehyde (2.4mg, 0.08mmol, 1.2eq), stirring at room temperature for 30min, adding sodium borohydride acetate (7mg, 0.033mmol, 0.5eq), stirring at room temperature for 2h, dropping saturated aqueous ammonium chloride solution for quenching, adding water (10mL) for dilution, ethyl acetate (20mL), concentrating the organic layer, washing with saturated salt water, drying with anhydrous sodium sulfate, concentrating under reduced pressure, and performing column chromatography to obtain 20mg of the product. ESI MS M/z 537[ M + H ]]⁺。

Example 11: synthesis of Compound 11

Intermediate 1(15mg, 0.029mmol, 1eq) was dissolved in THF (10mL), CDI (5.6mg, 0.034mmol, 1.2eq) was added, and after stirring at room temperature for 30min, 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (9.5mg, 0.034mmol, 1.2eq) was added, and after stirring at room temperature for 2h, the reaction was concentrated and purified by Pre-TLC to give 6mg of the product. ESI MS M/z 822[ M + H ]]⁺。

Example 12: synthesis of Compound 12

Intermediate 2(25mg, 0.051mmol, 1eq) was dissolved in THF (10mL), CDI (10mg, 0.06mmol, 1.2eq) was added, stirring was carried out at room temperature for 30min, 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (17mg, 0.06mmol, 1.2eq) was added, stirring was carried out at room temperature for 2h, the reaction was concentrated, and the product was purified by Pre-TLC to give 12 mg. ESI MS M/z 806[ M + H ]]⁺。

Example 13: synthesis of Compound 13

Intermediate 3(35mg, 0.069mmol, 1eq) was dissolved in THF (10mL) and CDI (13mg, 1eq) was added0.082mmol, 1.2eq), stirring at room temperature for 30min, adding 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (23mg, 0.083mmol, 1.2eq), stirring at room temperature for 2h, concentrating the reaction solution, and purifying by Pre-TLC to obtain 10mg of the product. ESI MS M/z 788[ M + H ]]⁺。

Example 14: synthesis of Compound 14

Intermediate 4(40mg, 0.080mmol, 1eq) was dissolved in THF (10mL), CDI (15mg, 0.095mmol, 1.2eq) was added, and after stirring at room temperature for 30min, 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (26mg, 0.095mmol, 1.2eq) was added, and after stirring at room temperature for 2h, the reaction was concentrated and Pre-TLC purified to give 12mg of product. ESI MS M/z 802[ M + H ] +.

Example 15: synthesis of Compound 15

Intermediate 5(41mg, 0.072mmol, 1eq) was dissolved in THF (10mL), CDI (14mg, 0.087mmol, 1.2eq) was added and stirred at room temperature for 30min, then 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (24mg, 0.087mmol, 1.2eq) was added and stirred at room temperature for 2h, then the reaction was concentrated and purified by Pre-TLC to give 13mg of product. ESI MS M/z 866[ M + H ] +.

Example 15: synthesis of Compound 15

Intermediate 6(20mg, 0.040mmol, 1eq) was dissolved in THF (10mL), CDI (7.8mg, 0.048mmol, 1.2eq) was added, stirring was carried out at room temperature for 30min, 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (13mg, 0.048mmol, 1.2eq) was added, stirring was carried out at room temperature for 2h, the reaction mixture was concentrated, and the product was purified by Pre-TLC to give 8mg of product。ESI MS m/z 796[M+H]⁺。

Example 17: synthesis of Compound 17

Intermediate 7(20mg, 0.042mmol, 1eq) was dissolved in THF (10mL), CDI (8.1mg, 0.050mmol, 1.2eq) was added, and after stirring at room temperature for 30min, 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (13.7mg, 0.050mmol, 1.2eq) was added, and after stirring at room temperature for 2h, the reaction was concentrated and purified by Pre-TLC to give 7mg of product. ESI MS M/z 777[ M + H ]]⁺。

Example 18: synthesis of Compound 18

Intermediate 8(15mg, 0.056mmol, 1eq) was dissolved in THF (10mL), CDI (6.1mg, 0.067mmol, 1.2eq) was added, stirring was carried out at room temperature for 30min, then 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (18mg, 0.067mmol, 1.2eq) was added, stirring was carried out at room temperature for 2h, then the reaction was concentrated, and the product was purified by Pre-TLC to give 5 mg. ESI MS M/z 778[ M + H ]]⁺。

Example 19: synthesis of Compound 19

Intermediate 9(25mg, 0.054mmol, 1eq) was dissolved in THF (10mL), CDI (10.4mg, 0.064mmol, 1.2eq) was added, stirring was carried out at room temperature for 30min, then 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (18mg, 0.064mmol, 1.2eq) was added, stirring was carried out at room temperature for 2h, then the reaction was concentrated, and the product was purified by Pre-TLC to give 7 mg. ESI MS M/z 766[ M + H ]]⁺。

Example 20: synthesis of Compound 20

Intermediate 1(15mg, 0.029mmol, 1eq) was dissolved in THF (10mL), CDI (5.6mg, 0.034mmol, 1.2eq) was added, and after stirring at room temperature for 30min, 3- (5- (methylaminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (9.9mg, 0.034mmol, 1.2eq) was added, and after stirring at room temperature for 2h, the reaction was concentrated and purified by Pre-TLC to give 7mg of the product. ESI MS M/z 836[ M + H ]]⁺。

Example 21: synthesis of Compound 21

Intermediate 10(20mg, 0.037mmol, 1eq) was dissolved in THF (10mL), CDI (7.2mg, 0.047mmol, 1.2eq) was added, and after stirring at room temperature for 30min, 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (12mg, 0.047mmol, 1.2eq) was added, and after stirring at room temperature for 2h, the reaction was concentrated and purified by Pre-TLC to give 8mg of product. ESI MS M/z 836[ M + H ]]⁺。

Example 22: synthesis of Compound 22

Intermediate 1(15mg, 0.029mmol, 1eq) was dissolved in THF (10mL), thiocarbonyldiimidazole (6mg, 0.034mmol, 1.2eq) was added, stirring was performed at room temperature for 30min, 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (9.5mg, 0.034mmol, 1.2eq) was added, stirring was performed at room temperature for 2h, the reaction was concentrated, and Pre-TLC purification gave 8mg of the product. ESI MS M/z 838[ M + H ]]⁺。

Example 23: synthesis of Compound 23

Intermediate 1(15mg, 0.029mmol, 1eq) was dissolved in THF (10mL), thiocarbonyldiimidazole (6.1mg, 0.034mmol, 1.2eq) was added and stirred at room temperature for 30min, then 3- (5- (methylaminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (9.9mg, 0.034mmol, 1.2eq) was added and stirred at room temperature for 2h, then the reaction was concentrated and purified by Pre-TLC to give 6mg of the product. ESI MS M/z 852[ M + H ]]⁺。

Example 24: synthesis of intermediate 24

4- ((1r,3r) -3-amino-2, 2,4, 4-tetramethylcyclobutylalkoxy) -2-chlorobenzonitrile (200mg, 0.72mmol, 1eq), 4-chloropyridazine-2-carboxylic acid (136mg, 0.86mmol, 1.2eq) were dissolved in DMF (15mL), DIEA (179. mu.L, 1.08mmol, 1.5eq) and HATU (410mg, 1.08mmol, 1.5eq) were added, respectively, and the mixture was stirred at room temperature for 2 h. After the TLC detection reaction, water (50mL) was added for dilution, ethyl acetate (50mL) was extracted three times, the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography to give 256mg of the product. The above product (50mg, 0.12mmol, 1eq) and methyl 3-chloro-4-aminobenzoate (24mg, 0.13mmol, 1.1eq) were dissolved in dioxane (15mL), HCl in dioxane (4M,5mL) was added dropwise, and the mixture was stirred at 80 ℃ overnight. After TLC detection, water (50mL) was added for dilution, NaHCO₃Adjusting the aqueous solution to neutrality, extracting with ethyl acetate (50mL), concentrating the organic layer, washing with saturated saline, drying with anhydrous sodium sulfate, concentrating under reduced pressure, and performing column chromatography to obtain 32 mg. The above product was dissolved in a mixed solvent of methanol and 10% NaOH solution (1:1,10mL) and stirred at room temperature for 2 h. After completion of the TLC detection reaction, the reaction mixture was diluted with water (20mL), washed with EA (20mL), the aqueous layer was adjusted to 4 with 2M hydrochloric acid, extracted with EA (20mL), the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 2515 mg of a product. ESI MS M/z 554[ M + H ]]⁺。

Example 25: synthesis of intermediate 25

Following the procedure of example 24, intermediate 2516 mg, ESI MS M/z 542[ M + H ]]⁺。

Example 26: synthesis of intermediate 26

Following the procedure of example 24, intermediate 2613 mg, ESI MS M/z 568[ M + H]⁺。

Example 27: synthesis of intermediate 27

Following the procedure of example 24, intermediate 2716 mg, ESI MS M/z 540[ M + H ]]⁺。

Example 28: synthesis of intermediate 28

Benzyl 4-iodobenzoate (200mg, 0.59mmol, 1eq), methyl 3-chloro-4-aminobenzoate (121mg, 0.65mmol, 1.1eq), palladium acetate (13mg, 0.059mmol, 0.1eq), BINAP (35mg, 0.059mmol, 0.1eq) and cesium carbonate (231mg, 0.65mmol, 1.1eq) were dissolved in dioxane (25mL) and stirred under nitrogen at 100 ℃ overnight. After TLC detection reaction, suction filtration, decompression concentration of filtrate and column chromatography are carried out to obtain 255mg of product. The above product was dissolved in THF (15mL), palladium hydroxide (25mg) was added, stirred overnight at room temperature under hydrogen, filtered, and the filtrate was concentrated to give 240mg of product, ESI MS M/z 306[ M + H ], (ESI MS M/z)]⁺. 4- ((1r,3r) -3-amino-2, 2,4, 4-tetramethylcyclobutylalkoxy) -2-chlorobenzonitrile (100mg, 0.36mmol, 1eq), 4-chloropyridazine-2-carboxylic acid (121mg, 0.43mmol, 1.2eq) were dissolved in DMF (15mL), DIEA (89. mu.L, 0.54mmol, 1.5eq) and HATU (205mg, 0.54mmol, 1.5eq) were added, respectively, and the mixture was stirred at room temperature for 2 h. TLC detection of reaction completionThen, water (50mL) was added for dilution, extraction was carried out with ethyl acetate (50mL) three times, the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography was carried out to give 82mg of the product. The above product was dissolved in a mixed solvent of methanol and 10% NaOH solution (1:1,10mL) and stirred at room temperature for 2 h. After completion of TLC detection, the reaction mixture was diluted with water (20mL), washed with EA (20mL), the aqueous layer was adjusted to 4 with 2M hydrochloric acid, extracted with EA (20mL), the organic layer was concentrated, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 2855 mg of a product. ESI MS M/z 552[ M + H ]]⁺。

Example 29: synthesis of intermediate 29

Following the procedure of example 28, intermediate 29, ESI MS M/z 518[ M + H ]]⁺。

Example 30: synthesis of intermediate 30

Following the procedure of example 28, intermediate 30, ESI MS M/z 536[ M + H]⁺。

Example 31: synthesis of intermediate 31

Following the procedure of example 28, intermediate 31, ESI MS M/z 532[ M + H ]]⁺。

Example 32: synthesis of intermediate 32

Following the procedure of example 28, intermediate 32, ESI MS M/z 548[ M + H ]]⁺。

Example 33: synthesis of intermediate 33

Following the procedure of example 28, intermediate 33, ESI MS M/z 519[ M + H ]]⁺。

Example 34: synthesis of intermediate 34

Following the procedure of example 28, intermediate 34, ESI MS M/z 519[ M + H ]]⁺。

Example 35: synthesis of intermediate 35

Following the procedure of example 28, intermediate 35, ESI MS M/z 566[ M + H ]]⁺。

Example 36: synthesis of intermediate 36

Following the procedure of example 28, intermediate 36, ESI MS M/z 532[ M + H ]]⁺。

Example 37: synthesis of intermediate 37

Following the procedure of example 28, intermediate 37, ESI MS M/z 550[ M + H ]]⁺。

Example 38: synthesis of intermediate 38

Following the procedure of example 28, intermediate 38, ESI MS M/z 546[ M + H]⁺。

Example 39: synthesis of intermediate 39

Following the procedure of example 28, intermediate 39, ESI MS M/z 533[ M + H ]]⁺。

Example 40: synthesis of intermediate 40

Following the procedure of example 28, intermediate 40, ESI MS M/z 533[ M + H ]]⁺。

Example 41: synthesis of Compound 41

Intermediate 24(15mg, 0.027mmol, 1eq), 3- (5- (aminomethyl) -1-oxoisoindolin-2-yl) piperidine-2, 6-dione (8.9mg, 0.033mmol, 1.2eq) were dissolved in DMF (15mL) and DIEA (5. mu.L, 0.033mmol, 1.2eq) and HATU (12mg, 0.033mmol, 1.2eq) were added separately and stirred at room temperature for 2 h. After the TLC detection reaction, the reaction mixture was concentrated under reduced pressure and then pre-TLC purified to obtain 10mg of a product. ESI MS M/z 809[ M + H ]]⁺。

Example 42: synthesis of Compound 42

Following the procedure of example 41, the product 42, ESI MS M/z 781[ M + H]⁺。

Example 43: synthesis of Compound 43

Following the procedure of example 41, product 43 was obtained, ESI MS M/z 823[ M + H ] +.

Example 44: synthesis of Compound 44

Following the procedure of example 41, the product 44 was obtained, ESI MS M/z 795[ M + H [ ]]⁺。

Example 45: synthesis of Compound 45

Following the procedure of example 41, product 45 was obtained, ESI MS M/z 807[ M + H [ ]]⁺。

Example 46: synthesis of Compound 46

Following the procedure of example 41, product 46 was obtained, ESI MS M/z 773[ M + H [ ]]⁺。

Example 47: synthesis of Compound 47

Following the procedure of example 41, product 47 was obtained, ESI MS M/z 791[ M + H [ ]]⁺。

Example 48: synthesis of Compound 48

Following the procedure of example 41, product 48, ESI MS M/z 787[ M + H ]]⁺。

Example 49: synthesis of Compound 49

Following the procedure of example 41, product 48 was obtained, ESI MS M/z 803[ M + H ]]⁺。

Example 50: synthesis of Compound 50

Following the procedure of example 41, product 50, ESI MS M/z 774[ M + H ]]⁺。

Example 51: synthesis of Compound 51

Following the procedure of example 41, product 51, ESI MS M/z 774[ M + H ]]⁺。

Example 52: synthesis of Compound 52

Following the procedure of example 41, product 52, ESI MS M/z 821[ M + H]⁺。

Example 53: synthesis of Compound 53

Following the procedure of example 41, product 53, ESI MS M/z 787[ M + H ]]⁺。

Example 54: synthesis of Compound 54

Following the procedure of example 41, the product 54, ESI MS M/z 805[ M + H ]]⁺。

Example 55: synthesis of Compound 55

Following the procedure of example 41, product 55 was obtained, ESI MS M/z 801[ M + H [ ]]⁺。

Example 56: synthesis of Compound 56

Following the procedure of example 41, product 56, ESI MS M/z 788[ M + H ]]⁺。

Example 57: synthesis of Compound 57

Following the procedure of example 41, the product 57 was obtained, ESI MS M/z 788[ M + H]⁺。

Effect example 1

Experimental methods

PC-3 cells: excell, cat # CL-0185. LNCaP cells: wuhan Punuo race life science and technology.

Antiproliferative activity of PC-3 cells

PC-3 cells were digested, counted, prepared into cell suspensions at a concentration of 2.2X 104 cells/mL, and 160. mu.L of cell suspension (3500 cells per well) was added to each well of a 96-well plate; culturing the 96-well plate in a 5% CO2 incubator at 37 ℃ for 24 hours; diluting the drug with complete culture medium to required concentration, and adding 40 μ L of corresponding drug-containing culture medium into each well; placing the 96-well plate in a 5% CO2 incubator at 37 ℃ for 4 d; finally, the cell activity was measured by the CCK-8 method, and the cell antiproliferative activity was calculated according to the formula ═ cell viability ((NC-blank) - (compound-blank))/((NC-blank) - (PC-blank)) × 100%. Nc (negative control) is the DMSO group and pc (positive control) is the drug with 100% lethality.

LNCap cell antiproliferative Activity

And (3) digesting and counting LNCap cells to prepare a cell suspension with the concentration of 3.13 multiplied by 104/mL, taking a sufficient volume of the cell suspension, adding a DCC culture medium to dilute the cell density to 5000 cells/160 uL, and mixing uniformly. 160uL of cell suspension is put into a 96-well plate, a PC group is added, then a proper amount of R1881 is added, 160uL of cell suspension is added into each hole, and the mixture is cultured in an incubator at 37 ℃ for 2 days. Diluting the drug with complete culture medium to required concentration, and adding 40 μ L of corresponding drug-containing culture medium into each well; placing the 96-well plate in a 5% CO2 incubator at 37 ℃ for 4 d; finally, the cell activity was measured by the CCK-8 method, and the cell anti-proliferative activity was calculated according to the formula ═ cell viability ((NC-blank) - (compound-blank))/((NC-blank) - (PC-blank)) × 100%. NC (negative control) is a control group to which R1881 (mettrenone) is added and PC (positive control) is a control group to which R1881 is added.

TABLE 1 inhibitory Activity of the Compounds of the present invention on LNCap cells and PC-3 cells

Claims

1. A compound represented by formula I or a pharmaceutically acceptable salt thereof;

wherein:

R¹is halogen, C₁-C₆Alkyl or C₁-C₆An alkoxy group;

R²is halogen, C₁-C₆Alkyl or C₁-C₆An alkoxy group;

x is O, S, CH₂Or NR³；R³Is hydrogen or C₁-C₆An alkyl group;

y is CH or N, R' is hydrogen or C₁-C₆An alkyl group;

n is 0 or 1, and when n is 0, Y is absent;

z is O or S;

r is hydrogen or C₁-C₆An alkyl group;

or the like, or, alternatively,r and R' are linked to form- (CH)₂)_m-structure, m is 2, 3 or 4.

2. The compound of formula I, or a pharmaceutically acceptable salt thereof, as claimed in claim 1, wherein

Is formed by one or more C₁-C₆Alkyl substituted C₄-C₇When cycloalkylene, said C₁-C₆Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

and/or when

Is formed by one or more C₁-C₆Alkyl substituted C₄-C₇When cycloalkylene, said C₄-C₇Cycloalkylene is cyclobutyl, cyclopentyl, cyclohexylene or cycloheptylene;

and/or when

Is formed by one or more C₁-C₆Alkyl substituted C₄-C₇When cycloalkylene, said plurality is two, three or four;

and/or when

Is C₄-C₇When cycloalkylene, said C₄-C₇Cycloalkylene is cyclobutyl, cyclopentyl, cyclohexylene or cycloheptylene;

and/or when

When it is phenylene, said phenylene is

And/or when

In the case of "a 5-6 membered heteroarylene group having 1,2 or 3 hetero atoms independently selected from one or more of N, O and S", the 5-6 membered heteroarylene group is a triazolylene group, a pyrazolyl arylene group, a pyridyl ene group, a pyrimidylene group, a pyrazinylene group, an oxazolylene group or a pyridazinylene group;

and/or when

When it is phenylene, said phenylene is

And/or when R¹When the halogen is fluorine, chlorine, bromine or iodine;

and/or when R¹Is C₁-C₆When alkyl, said C₁-C₆Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

and/or when R¹Is C₁-C₆At alkoxy, said C₁-C₆Alkoxy is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy;

and/or when

Is represented by one or more R¹When substituted phenylene, said phenylene is

And/or when

Is represented by one or more R¹When substituted phenylene, said plurality is two or three;

and/or when

In the case of "a 5-6 membered heteroarylene group having 1,2 or 3 hetero atoms independently selected from one or more of N, O and S", the 5-6 membered heteroarylene group is a triazolylene group, a pyrazolyl arylene group, a pyridyl ene group, a pyrimidylene group, a pyrazinylene group, an oxazolylene group and a pyridazinylene group;

and/or when R²When the halogen is fluorine, chlorine, bromine or iodine;

and/or when R²Is C₁-C₆When alkyl, said C₁-C₆Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

and/or when R²Is C₁-C₆At alkoxy, said C₁-C₆Alkoxy is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy;

and/or when

Is "by one or more R²(ii) when the number of substituted atoms is 1,2 or 3 and the heteroatom is independently selected from N, O and S, then the 5-6 membered heteroarylene group is triazolylene, pyrazolyl, pyridylene, pyrimidinyl, pyrazinylene, oxazolylene or pyridazinylene;

and/or when R³Is C₁-C₆When alkyl, said C₁-C₆Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

and/or, when R' is C₁-C₆When alkyl, said C₁-C₆Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

and/or, when R is C₁-C₆When alkyl, said C₁-C₆Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

3. The compound of formula I, or a pharmaceutically acceptable salt thereof, as claimed in claim 2, wherein

Is formed by one or more C₁-C₆Alkyl substituted C₄-C₇When cycloalkylene, said C₁-C₆Alkyl is methyl;

and/or when

Is formed by one or more C₁-C₆Alkyl substituted C₄-C₇When cycloalkylene, said C₄-C₇Cycloalkylene is

Preference is given to

And/or when

Is C₄-C₇When cycloalkylene, said C₄-C₇Cycloalkylene is

Preference is given to

And/or when

When it is phenylene, said phenylene is

And/or when

When the "5-6 membered heteroarylene group having 1,2 or 3 hetero atoms and one or more hetero atoms independently selected from N, O and S" is used, the 5-6 membered heteroarylene group is

And/or when

When it is phenylene, said phenylene is

And/or when R¹When it is halogen, said halogen is fluorine, chlorine orBromine;

and/or when R¹Is C₁-C₆When alkyl, said C₁-C₆Alkyl is methyl;

and/or when R¹Is C₁-C₆At alkoxy, said C₁-C₆Alkoxy is methoxy;

and/or when

Is represented by one or more R¹When substituted phenylene, said phenylene is

Said quilt is an R¹Substituted phenylene radicals are preferred

It is also preferable that

And/or, when

And/or when R²When the halogen is fluorine, chlorine or bromine;

and/orWhen R is²Is C₁-C₆When alkyl, said C₁-C₆Alkyl is methyl;

and/or when R²Is C₁-C₆At alkoxy, said C₁-C₆Alkoxy is methoxy;

and/or when

Is "by one or more R²When the substituted 5-6 membered heteroarylene group has 1,2 or 3 substituted atoms and the hetero atom is independently selected from N, O and S, the 5-6 membered heteroarylene group is

And/or when R³Is C₁-C₆When alkyl, said C₁-C₆Alkyl is methyl;

and/or, when R' is C₁-C₆When alkyl, said C₁-C₆Alkyl is methyl;

and/or, when R is C₁-C₆When alkyl, said C₁-C₆The alkyl group is a methyl group.

4. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 2,

And/or the presence of a gas in the gas,

is composed of

And/or the presence of a gas in the gas,

is phenylene, substituted by an R¹Substituted phenylene or "5-6 membered heteroarylene having 1,2 or 3 heteroatoms independently selected from one or more of N, O and S", preferably

5. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1,

And/or the presence of a gas in the gas,

is phenylene, preferably

And/or the presence of a gas in the gas,

is phenylene or "substituted by one or more R¹Substituted phenylene radicals ";

and/or, R¹Is hydrogen;

and/or, R' is hydrogen;

and/or n is 1;

and/or, R is hydrogen;

preferably, the first and second liquid crystal films are made of a polymer,

is composed of

6. The compound of formula I, or a pharmaceutically acceptable salt thereof, according to claim 1, wherein the compound of formula I is according to any one of the following schemes:

the first scheme is as follows:

Is phenylene, the number of "hetero atoms" is 1,2 or 3, the hetero atoms being independentOne or more 5-6 membered heteroarylene groups selected from N, O and S, said 5-6 membered heteroarylene group being a pyridazinylene group, preferably

The 5-to 6-membered heteroarylene group is preferred

Scheme II:

The third scheme is as follows:

Is phenylene, preferably

R¹Is halogen or C₁-C₆An alkyl group;

n is 1;

y is N and R' is hydrogen.

7. The compound of formula I, or a pharmaceutically acceptable salt thereof, according to claim 1, wherein the compound of formula I is any one of the following compounds:

8. a method for preparing a compound of formula I according to any one of claims 1 to 7, characterized in that it is any one of the following schemes:

scheme A: when n is 0, or n is 1 and Y is CH, the preparation method of the compound shown in the formula I is as follows:

scheme B: when N is 1 and Y is N, the preparation method of the compound shown in the formula I is as follows:

is as defined in any one of claims 1 to 6, X is as defined in any one of claims 1 to 3, R' is as defined in any one of claims 1 to 3 or 5, Y is as defined in claim 1 or 6, n is as defined in any one of claims 1, 5 or 6, and R is as defined in any one of claims 1 to 3 or 5.

9. A pharmaceutical composition, which comprises a substance X and a pharmaceutical adjuvant, wherein the substance X is a compound as shown in formula I or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 7, and the dosage of the substance X is preferably a therapeutically effective amount.

10. Use of substance X, or a pharmaceutical composition according to claim 9, in the preparation of an LNCap cell and/or PC-3 cell proliferation inhibitor, said substance X being a compound according to formula I, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 7.

11. Use of substance X, or a pharmaceutical composition according to claim 9, for the manufacture of a medicament for the treatment and/or prevention of a disorder in which the androgen receptor is involved, preferably a cancer, preferably prostate cancer, substance X being a compound according to formula I, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 7.