WO2019015689A1 - Chiral 3-quinuclidone compound, preparation method, and use - Google Patents

Abstract

Provided are a chiral 3-quinuclidone compound, a preparation method, and a use. In particular, the present invention provides a compound shown in general formula I-A or I-B or a pharmaceutically acceptable salt, isomer or a mixture thereof, solvate, polymorph, stable isotopic derivative, or prodrug thereof and a preparation method and application of the same. The compound has good cell proliferation inhibitory activity and pharmacokinetic properties and can be used to treat cancer, autoimmune diseases, etc. (IA) (IB)

Description

Chiral 3-quinuclidinone compound, preparation method and use thereof

This application claims priority from Chinese Patent Application No. CN201710603967.2, filed on July 21, 2017. This application cites the entire text of the above-mentioned Chinese patent application.

Technical field

The present invention relates to a chiral 3-quinuclidinone compound, an isomer thereof, a prodrug, a pharmaceutically acceptable salt, a stable isotope derivative, a pharmaceutical composition, a preparation method and use.

Background technique

High mortality rates in cancer and resistance to conventional cancer therapies remain a major challenge for human health. The current new therapeutic strategy focuses on the discovery of new non-chemotherapy targets for cancer-specific targets. The p53 gene is an important tumor suppressor gene that plays a key role in cell cycle regulation, DNA repair, apoptosis, etc. Mutant p53 gene is found in at least 50% of tumors, and most of the chemotherapeutic drugs that induce DNA damage require functional p53 to exert its anti-tumor effect, while p53 mutation or indirect mechanism causes inactivation of p53 to increase resistance of traditional drugs. Sexuality, treatment with tumors carrying mutant p53 genes is currently a major challenge, and thus p53 has become an important target gene for antitumor drugs.

There are two types of p53 gene: wild type and mutant type. Wild type p53 is an important tumor suppressor gene in human body, but the mutant p53 has a conformational change and loses its tumor suppressor function, and also inhibits the transcriptional activity of wild-type p53. Loss of inhibition of cell growth, causing abnormal cell proliferation and transformation into malignant cells. Currently, malignant tumors closely related to p53 gene mutations have liver cancer, stomach cancer, colon cancer, esophageal cancer, lung cancer, breast cancer, lymphoma, ovary Cancer, cervical cancer, etc. At present, p53-targeted drugs are mainly used in the following ways: gene therapy, such as the drugs Gendicine and Advexin; second, the mechanism of action of p53-MDM2, which prevents p53 by interfering with the synthesis of p53-MDM2 complex by small molecule drugs. Degradation of protein ubiquitination, such as drugs Idasanutlin, HDM-201 and AMG-232; third is the change of conformation of mutant p53 protein, small molecule compound binds to amino acid by covalent bond, thereby changing or modifying the structure of p53 To change its conformation, thereby restoring the activity of the tumor suppressor gene, such as the compound APR-246/PRIMA-1 ^MET , which is metabolized in the body to methylenequinucdinone MQ, which becomes the Michael receptor through cysteine. The thiol group of the protein undergoes additive binding to form a covalent bond-modified DNA, which changes the conformation of the p53 protein and activates transcriptional activity. Currently, the compound enters a clinical phase II study for the treatment of esophageal cancer, prostate cancer, and acute myeloid leukemia. , ovarian cancer and myeloproliferative tumors.

APR-246 is a clinically staged treatment of racemic 3-quinuclidinone compounds that carry mutant p53 gene tumors, while the enantiomers of drugs often have different biological activities, pharmacokinetic properties, Toxic side effects and metabolic stability, it is therefore important and necessary to develop optically active 3-quinuclidinone compounds.

Summary of the invention

It is an object of the present invention to provide a class of chiral 3-quinuclidinone compounds, processes for their preparation and uses. The 3-quinuclidinone compound of the present invention has good cell proliferation inhibitory activity and pharmacokinetic properties.

In a first aspect, the present invention provides a compound represented by the formula IA or IB, which (refers to a compound of the formula IA or IB), a pharmaceutically acceptable salt, an isomer or a mixture thereof, a solvate thereof , a polymorph, a stable isotope derivative or a prodrug;

among them,

R is selected from substituted or unsubstituted, unbranched or branched, saturated or unsaturated C _1-10 alkyl groups (such as C _1-6 alkyl groups, such as C _2-6 alkyl groups, such as C _{2 -4} alkyl) or C _3-12 cycloalkyl (such as C _3-6 cycloalkyl); substituted or unsubstituted aryl (such as C _6-12 aryl); substituted or unsubstituted saturated or unsaturated Heterocycle (such as a C _5-7 heterocycle, wherein the number of heteroatoms may be one or more, such as 1, 2 or 3; each heteroatom may be selected from N, O or S); -L ¹ -L ² , L ^{1 is} selected from a substituted or unsubstituted C _1-10 alkyl group (such as a C _1-4 alkyl group, such as a methyl group), and L ^{2 is} selected from an aryl group (such as a C _6-12 aryl group). a heteroaryl group (such as a C _6-12 heteroaryl group, the number of which may be one or more, such as 1, 2 or 3; each hetero atom may be selected from N, O or S) or a ring (such as a C _5-7 heterocyclic ring, the number of which may be one or more, such as 1, 2 or 3; each hetero atom may be selected from N, O or S), in the heterocyclic ring The hetero atom is selected from N, O, S or P.

In a preferred embodiment of the invention, R is selected from substituted or unsubstituted C _1-10 alkyl, preferably substituted or unsubstituted C _1-6 alkyl, such as methyl, trifluoromethyl, ethyl, propyl Base, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

In a preferred embodiment of the invention, R is selected from -L ¹ -L ² , L ^{1 is} selected from substituted or unsubstituted C _1-6 alkyl, more preferably C _1-4 alkyl; L ² is preferably derived from aryl or Heterocycle, such as benzyl, phenylethyl, phenylpropyl or phenylbutyl.

In a preferred embodiment of the invention, R is selected from the group consisting of methyl.

In a preferred embodiment of the present invention, the compound represented by Formula I-A or I-B is selected from any of the following structures:

In some embodiments of the invention, the compound of Formula I can be separately collected under chiral resolution conditions at RT for 12.2 minutes or 14.0 minutes;

Compound 1

Or, the compound of formula I can be separately collected under chiral resolution conditions at an RT of 13.2 minutes or 15.4 minutes;

Compound 2

Or, the compound of formula I can be separately collected under chiral resolution conditions at an RT of 15.2 minutes or 16.9 minutes;

Compound 3

Or, a compound of the formula I can be separately collected under chiral resolution conditions at an RT of 14.8 minutes or 17.8 minutes;

Compound 4

Or, the compound of formula I can be separately collected under chiral resolution conditions at RT for 12.5 minutes or 14.4 minutes;

Compound 5

Or, the compound of Formula I can be separately collected under chiral resolution conditions at an RT of 14.3 minutes or 16.2 minutes;

Compound 6

Or, a compound of the formula I can be obtained by separately collecting the compound 7 at an RT of 12.9 minutes or 14.6 minutes;

Compound 7

Or, a compound of the formula I can be obtained by separately collecting the compound 8 at an RT of 10.5 minutes or 12.1 minutes;

Compound 8

Or, the compound of formula I can be obtained by separately collecting compound 9 at 8.9 minutes or 10.7 minutes at RT;

Compound 9

The chiral splitting conditions may include:

The chiral column is CHIRALPAK AS-H 10mm x 250mm, 5μm;

The column temperature is 40 ° C;

Mobile phase A is 0.1% DEA in Hexane, and the percent is volume percent;

Mobile phase B is ethanol;

The gradient is mobile phase A / mobile phase B = 90/10, the ratio is the volume ratio;

The flow rate is 6.0 mL/min;

The detection wavelength was UV 310 nm.

Even if other chiral resolution, purification methods or detection methods are employed, it is within the scope of the present invention to obtain a chiral compound at a corresponding retention time under the chiral separation or detection method described in the present invention.

In a second aspect, the present invention provides a compound of the formula IA or IB, a pharmaceutically acceptable salt, an isomer thereof or a mixture thereof, a solvate, a polymorph, a stable isotope A method for preparing a derivative or a prodrug, comprising the steps of:

The 3-quinuclidinone is reacted with formaldehyde and an alcohol ROH under basic conditions to obtain a racemic compound of the formula IC, and the compound of the formula IC can be subjected to a chiral column or chemical resolution. Sexually isolated to give compounds as shown in Formulas IA and IB;

Wherein the definition of the R group is as described above.

In the preparation method, in the reaction for preparing a compound represented by the formula I-C, the reaction solvent may be a conventional solvent for such a reaction in the art, and for example, the reaction solvent may be a mixed solvent of an alcohol ROH and water. The volume ratio of the alcohol ROH to water may be a conventional volume ratio of such a reaction in the art, for example, from 3:1 to 1:2.

In the preparation method, in the reaction for preparing a compound represented by the formula IC, the 3-quinuclidinone may be added as an inorganic salt of 3-quinuclidinone (such as 3-quinuclidinone hydrochloride). salt).

In the preparation method, in the reaction of preparing a compound represented by the formula IC, the amount of formaldehyde may be a conventional amount for such a reaction in the field, for example, the molar ratio of formaldehyde to 3-quinuclidinone may be 5-20. :1.

In the preparation method, the reagent for providing basic conditions may be a conventional alkaline reagent for the reaction in the field, including an organic base and an inorganic base, and the organic base may be lithium hexamethyldisilazide. , hexamethyldisilazide sodium, hexamethyldisilazide potassium, diisopropylamide lithium, n-butyllithium, sec-butyllithium, triethylamine, pyridine, 2,6-lutidine One or more of N,N-diisopropylethylamine, potassium t-butoxide, sodium t-butoxide, lithium t-butoxide, tetrabutylammonium fluoride and N-methylmorpholine. The inorganic base may be one of potassium carbonate, sodium carbonate, sodium hydrogencarbonate, sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium fluoride, cesium carbonate, lithium carbonate, potassium phosphate, sodium hydride and potassium hydride. Kind or more. For example, the alkaline agent can be potassium carbonate. The amount of the alkaline agent used may be a conventional amount for such a reaction in the art, and for example, the molar ratio of the alkaline agent to 3-quinuclidinone may be from 1 to 10:1.

In the preparation method, in the reaction for preparing a compound represented by the formula IC, the reaction temperature may be a usual reaction temperature for such a reaction in the art, such as -78 ° C - 250 ° C (eg, 30-90 ° C, and 60). -80 ° C).

In the preparation method, the progress of the reaction for preparing a compound such as the compound of the formula IC can be monitored by a conventional test method (such as TLC, HPLC, GC or NMR) in the art, generally 3-pyrininone. The reaction was carried out as the end point of the reaction.

In the preparation method, the reaction for preparing the compound represented by the formula I-C may further comprise the following post-treatment step: after the reaction is completed, the aqueous solution of sodium hydroxide is added to adjust the pH to 12-13, extracted, concentrated, and purified.

In one embodiment of the invention, the reaction for preparing a compound of the formula IC includes the steps of mixing 3-quinuclidinone hydrochloride, aqueous formaldehyde solution and potassium carbonate in an alcohol ROH and water at 60-80. After the reaction is completed, an aqueous sodium hydroxide solution is added to adjust the pH to 12-13, extracted, concentrated, and separated and purified.

In the preparation method, the chiral column may be a conventional chiral column in the art, such as CHIRALPAK AS-H. The chiral resolution conditions may employ conventional chiral resolution conditions in the art, including, for example, the following conditions:

The chiral column is CHIRALPAK AS-H 10mm x 250mm, 5μm;

The column temperature is 40 ° C;

Mobile phase A is 0.1% DEA in Hexane, and the percent is volume percent;

Mobile phase B is ethanol;

The gradient is mobile phase A / mobile phase B = 90/10, the ratio is the volume ratio;

The flow rate is 6.0 mL/min;

The detection wavelength was UV 310 nm.

In a third aspect, the present invention provides a pharmaceutical composition comprising an effective amount of a compound represented by Formula IA or IB as described above, a pharmaceutically acceptable salt thereof, an isomer thereof, or a mixture thereof, a solvate thereof. a polymorph, a stable isotope derivative or prodrug, and a pharmaceutically acceptable carrier, diluent or excipient;

In a fourth aspect, the present invention provides a compound of the formula IA or IB as described above, a pharmaceutically acceptable salt, an isomer thereof or a mixture thereof, a solvate, a polymorph, a stable isotope Use of a derivative or prodrug, or a pharmaceutical composition as described, for the manufacture of a medicament for the prevention, alleviation and/or treatment of a p53 mutation-related disease.

The p53 mutation-related disease may be selected from cancer, inflammatory disease, autoimmune disease, cardiovascular disease, heart disease or viral infection.

The cancer may be selected from the group consisting of ovarian cancer, prostate cancer, myeloma (such as myelodysplastic), esophageal cancer, gastric cancer, acute myeloid leukemia, chronic myelomonocytic leukemia, acute or chronic leukemia, and the like.

In a fifth aspect, the present invention provides a compound represented by the formula IA or IB, a pharmaceutically acceptable salt, an isomer thereof or a mixture thereof, a solvate, a polymorph, a stable isotope derivative Or prodrug, or the use of the pharmaceutical composition in combination with another anti-cancer agent.

In a sixth aspect, the present invention provides a compound of the formula IA or IB, a pharmaceutically acceptable salt, an isomer thereof or a mixture thereof, a solvate, a polymorph, a stable isotope derivative Or a prodrug, or the use of the pharmaceutical composition for the preparation of a medicament for treating cancer, wherein the medicament can be used in combination with another anti-cancer agent.

The anticancer agent may be selected from an alkylating agent (such as cyclophosphamide, hydrochloric acid mustard, dibromomannitol, carmustine, dacarbazine, melphalan, etc.), a platinum drug (such as cisplatin, Carboplatin, thioplatinum, nedaplatin, oxaliplatin, lobaplatin, etc., topoisomerase inhibitors (such as topotecan, irinotecan, rubatecan, exenatide, letote Kang, jimatidine, difluentine), DNA/RNA synthesis inhibitors (such as 5-Fluoracil, 5-FU, etc.), DNA methyltransferases (such as Azacitidine, Decitabine, etc.), liposomes (such as PLD) Etc., metabolic antagonists (such as methotrexate, capecitabine, pemetrexed, etc.), alkaloids (such as docetaxel, paclitaxel, vinblastine, etc.), antibody drugs (such as trastuzumab, Patricidumab, bevacizumab, etc.), hormone anticancer agents (such as leuprolide, dutasteride, dexamethasone, etc.), proteasome inhibitors (such as boraxomomi, azazomi, Lenalidomide, etc.), CDK kinase inhibitors (such as palbociclib, ribociclib, etc.), VEGFR or EGFR inhibitors (such as afatinib, imatinib, gefitinib, erlotinib, etc.), m- TOR inhibitors (eg everolimus, sirolimus) Etc.), PI3K kinase inhibitors (such as elialix, etc.), B-Raf inhibitors (such as sorafenib, vemurafenib, revafenib, etc.), other PARP inhibitors (such as olaparib, niraparib, etc.) ), HDAC inhibitors (such as citabin, paclitaxel, vorinostat, etc.) c-Met kinase inhibitors (such as crizotinib), ALK kinase inhibitors (such as ceritinib, alexidine) Nis et al), AKT inhibitors (such as piperfine, etc.), ABL inhibitors, FLT3 inhibitors, PD-1 monoclonal antibodies (such as Opdivo, Keytruda, etc.), PD-L1 monoclonal antibodies (such as Atezolizumab) and the like.

Detailed description of the invention

Unless stated to the contrary, the following terms used in the specification and claims have the following meanings:

The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight or branched chain groups of 1 to 20 carbon atoms. An alkyl group of 1 to 10 carbon atoms is preferred, more preferably 1 to 8 carbon atoms, and non-limiting examples include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, isobutyl, t-butyl , sec-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethyl Propyl, 1-ethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1,1-dimethylbutyl, 1,2-dimethyl Butyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1,1,2- Trimethylpropyl, 1-ethyl-2-methylpropyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,2- Dimethylpentyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 3,3-dimethylpentyl, 3,4-dimethylpentyl, 2-ethyl Butyl, 3-ethylpentyl, sulfhydryl, decyl, decyl, undecyl, dodecyl, and various isomers thereof and the like. The alkyl group may be substituted or unsubstituted, and when substituted, may be substituted at any available point of attachment, preferably one or more groups, independently selected from alkyl, halo, hydroxy. , mercapto, cyano, alkenyl, alkynyl, alkoxy, alkanoyl, alkylamino, nitro, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkane An oxy group, a cycloalkyl fluorenyl group, a heterocycloalkyl fluorenyl group, an oxo group, an amino group, a halogenated alkyl group, a hydroxyalkyl group, a carboxyl group or a carboxylate group. When "alkyl" and its prefix are used herein, both straight-chain and branched saturated carbon bonds are included.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic substituent comprising from 3 to 20 carbon atoms, preferably from 3 to 12 carbon atoms, more preferably from 3 to 10 carbon atoms, most preferably Non-limiting examples of including 3 to 6 carbon atoms, monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexyl Dienyl, cycloheptyl, cyclooctyl and the like. Non-limiting examples of polycyclic cycloalkyl groups include, but are not limited to, spiro, fused, and bridged cycloalkyl groups. The cycloalkyl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more groups independently selected from the group consisting of alkyl, halogen, hydroxy, decyl, cyano, alkenyl, alkyne. Alkyl, alkoxy, alkyl sulfonyl, alkylamino, nitro, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkyl decyl, heterocycloalkane Anthracenyl, oxo, amino, haloalkyl, hydroxyalkyl, carboxy or carboxylate groups, and the like.

The term "aryl" refers to any stable conjugated hydrocarbon ring system group of 6 to 18 carbon atoms, preferably 6 to 10 carbon atoms, which may be a monocyclic, bicyclic, tricyclic or more cyclic aromatic group. Groups such as phenyl, naphthyl and anthracene, etc., which may be fused to a heteroaryl, heterocycloalkyl or cycloalkyl ring. The aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more groups independently selected from the group consisting of alkyl, halogen, hydroxy, decyl, cyano, alkenyl, alkynyl, Alkoxy, alkylalkyl, alkylamino, nitro, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkyl fluorenyl, heterocycloalkyl fluorenyl, An oxo group, an amino group, a halogenated alkyl group, a hydroxyalkyl group, a carboxyl group or a carboxylate group, and the like.

The term "heterocycle" refers to a monocyclic, bicyclic, tricyclic or tetracyclic ring system, including fused ring systems, bridged ring systems or spiro ring systems, wherein one or more atoms on the ring are independently, optionally, replaced by a hetero atom, The hetero atom is selected from the group consisting of nitrogen, oxygen, sulfur, phosphorus and boron, etc., and the ring may be fully saturated or contain one or more unsaturations, but non-aromatic; nitrogen, carbon, phosphorus or sulfur in the heterocyclic group The atom may be optionally oxidized; the nitrogen atom may optionally be quaternized; the heterocyclic ring system may be attached to the main structure at any heteroatom or carbon atom to form a stable compound. The hydrogen atoms on one or more of the rings are independently, optionally, substituted by one or more substituents described herein. Examples of heterocyclic rings include, but are not limited to, pyrrolidinyl, pyrazolidinyl, morpholinyl, piperazinyl, piperidinyl, thiomorpholinyl, pyranyl, tetrahydropyranyl, imidazolidinyl, Indoline, octahydroindenyl, octahydroisodecyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, azetidinyl and the like.

The term "heteroaryl" refers to an aromatic ring system formed by the replacement of a carbon atom on at least one ring with a heteroatom selected from N, O or S, preferably a 5 to 7 membered monocyclic structure or a 7 to 12 membered bicyclic structure. More preferably, it is a 5- to 6-membered heteroaryl group such as pyrrolyl, imidazolyl, pyridyl, pyrimidinyl, thiazolyl, thienyl, pyrazinyl, triazolyl, tetrazolyl, oxazolyl, oxazolyl. Etc., the heteroaryl ring can be fused to an aryl, heterocycloalkyl or cycloalkyl ring. The heteroaryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more groups independently selected from the group consisting of alkyl, halogen, hydroxy, decyl, cyano, alkenyl, alkynyl. , alkoxy, alkylalkyl, alkylamino, nitro, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkyl fluorenyl, heterocycloalkyl fluorenyl An oxo group, an amino group, a halogenated alkyl group, a hydroxyalkyl group, a carboxyl group or a carboxylate group.

"Substituted" means that one or more hydrogen or deuterium atoms in the group, preferably from 1 to 5 hydrogen or deuterium atoms, are independently substituted with each other by a corresponding number of substituents.

"Pharmaceutically acceptable salt" refers to a biological group capable of retaining the free base without other toxic side effects, which may be an acidic group, a basic group or an amphoteric group, and non-limiting examples include, but are not limited to: Acid salts include hydrochloride, hydrobromide, sulfate, pyrosulfate, hydrogen sulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, coke Phosphate, nitrate, acetate, propionate, citrate, octoate, formate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate , succinate, suberate, benzoate, methyl benzoate, phthalate, maleate, methanesulfonate, p-toluenesulfonate, besylate, (D,L)-tartaric acid, citrate, maleate, (D, L-) malate, fumarate, stearate, oleate, cinnamate, laurate, Glutamate, aspartate, triflate, tonsilate, ascorbate, salicylate, and the like. When the compound of the present invention contains an acidic group, the pharmaceutically acceptable salt thereof may further include an alkali metal salt (for example, a sodium salt or a potassium salt), an alkaline earth metal salt (for example, a calcium salt or a magnesium salt), and an organic base salt (for example). Alkyl aromatic amino acids, amino acids, etc.).

"Solvate" means an aggregate (or associate) formed by one or more solvent molecules with a compound of the invention. Solvents of the solvate formed include, but are not limited to, water, dimethyl sulfoxide, methanol, ethanol, isopropanol, acetic acid, and the like.

"Polymorph" refers to a different solid crystalline phase of a compound of the invention produced in the solid state by the presence of two or more different molecular arrangements, which may exist as a single crystalline or polymorphic mixture.

The "stable isotope derivative" means an isotope-substituted derivative obtained by substituting a hydrogen atom of any one of the compounds of the present invention with 1 to 5 atomic atoms, or an isotope obtained by substituting one or three C ¹⁴ atoms for any carbon atom. An isotopic derivative obtained by substituting a derivative or an arbitrary oxygen atom with 1 to 3 O ¹⁸ atoms.

"Prodrug" means a compound that can be converted to a biologically active compound of the invention under physiological conditions (eg, in vivo) or by solvolysis, and is understood to be a pharmaceutically acceptable metabolic precursor. The prodrug may be inactive or less active than the active parent compound, but may be rapidly converted in vivo to produce the parent compound of the invention, which may improve its solubility in animals as well as certain metabolic properties, including, for example, amino protecting groups, A carboxyl protecting group, a phospholipid, or the like.

"Pharmaceutical composition" means a mixture comprising one or more of the compounds described herein, or a physiologically pharmaceutically acceptable salt or prodrug thereof, and other chemical components, as well as other components such as physiologically pharmaceutically acceptable carriers and excipient. The purpose of the pharmaceutical composition is to promote the administration of the organism, and to facilitate the absorption of the active ingredient to exert biological activity.

"Isomer" means a stereoisomer comprising: an enantiomer and a diastereomer, and a cis-trans isomer is one of the diastereomers. The isomers of the present compounds may be their enantiomers, diastereomers, and any mixture thereof, including the formation of free or salt forms.

The term "effective amount" or "therapeutically effective amount" refers to a sufficient amount of a drug or agent that is non-toxic but that achieves the desired effect. The determination of the effective amount will vary from person to person, depending on the age and general condition of the recipient, and also on the particular active substance, and a suitable effective amount in a case can be determined by one skilled in the art based on routine experimentation.

Abbreviations for any protecting groups, amino acids, and other compounds used in the present invention, unless otherwise indicated, are based on their commonly accepted and accepted abbreviations, or reference to the IUPAC-IUBC Commission on Biochemical Nomenclature (see Biochem. 1972, 11,942-944).

The above preferred conditions can be arbitrarily combined without departing from the ordinary knowledge in the art, that is, preferred embodiments of the present invention.

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

The positive progress of the present invention is that the 3-quinuclidinone compound of the present invention has better cell proliferation inhibiting activity and pharmacokinetic properties.

Detailed ways

The description is further described by the following examples, but these examples are not intended to limit the scope of the invention.

The experimental methods in the examples of the present invention which do not specify the specific conditions are generally in accordance with conventional methods and conditions, or in accordance with the conditions recommended by the raw material or commodity manufacturer. Reagents without specific source are routine reagents purchased from the market.

The structure of all compounds of the invention can be determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). The NMR shift (δ) is ^reported in units of 10 ^-6 (ppm). The NMR measuring instrument was performed on a Bruker AVANCE-400 spectrometer. The deuterated solvents tested were deuterated chloroform (CDCl ₃ ), deuterated methanol (MeOD), deuterated dimethyl sulfoxide (DMSO-D ₆ ), and the internal standard was tetramethylsilane (TMS).

Low resolution mass spectra (MS) were determined by an Agilent 6120 quadruple LCMS mass spectrometer.

HPLC purity was determined by Agilent HPLC 12000/1220 (Agilent Zorbax Bonus RP 3.5 μm × 4.6 mm × 150 mm or Boston pHlex ODS 4.6 mm × 150 mm × 3 μm or Sepax BR-C18 (4.6 mm x 150 mm) X3μm)

Purification of the compounds of the invention and their intermediates can be carried out using conventional preparative HPLC, silica gel plates, column chromatography or separation using a flash separator.

The thin layer chromatography silica gel plate uses Yantai Huanghai, Yantai Xinnuo Chemical HSGF254 or Qingdao GF254 silica gel plate. The silica gel plate used for thin layer chromatography (TLC) is 2.5x 5cm, 0.2mm～0.25mm, thin layer. The pre-TLC purification product has a specification of 1 mm or 0.4 mm to 0.5 mm and 20 x 20 cm.

Column chromatography (silica gel column chromatography) generally uses a specification of 100 to 200 mesh or 200 to 300 mesh or 300 to 400 mesh.

The instrument used in the fast separator is the Agela Technologies MP200, and the column size is generally Flash column silica-CS.

The instrument used for preparative HPLC (Pre-HPLC) was Gilson GX-281, column model: Welch ultimate XB-C18 21.2 mm x 250 mm x 10 μm.

The chiral test column models are CHIRALCEL OD-H, OJ-H or CHIRALPAK AD-H, AS-H 4.6mm x 250mm x 5μm, preparative column type CHIRALCEL OD-H, OJ-H or CHIRALPAK AD-H, AS -H 10mm x 250mm x 5μm.

Known starting materials of the present invention can be synthesized by or according to methods known in the art, or from suppliers Sigma Aldrich, ACROS, Alfa Aesar, TCI, Belling, Anne Chemical, Suiyuan Chemical, Maclean, Companies such as Siyan Chemical purchased the proceeds.

Anhydrous solvents such as anhydrous tetrahydrofuran, anhydrous dichloromethane, anhydrous N,N-dimethylacetamide and the like are all purchased from the above chemical companies.

Unless otherwise specified in the examples, the reaction is generally carried out under a nitrogen or argon atmosphere. The nitrogen or argon atmosphere means that the reaction flask is connected to a balloon of nitrogen or argon having a volume of about 1 L and subjected to three pumping displacements.

There is no particular description in the examples, the reaction temperature is room temperature, and the temperature is 15 to 25 °C.

The reaction in the examples is generally monitored by LCMS or TLC. The LCMS instrument is described above. The developing solvent system used in TLC is generally: dichloromethane and methanol, petroleum ether and ethyl acetate, dichloromethane and acetic acid. Ester, petroleum ether and system of dichloromethane, ethyl acetate and methanol, the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount (0.1% to 10%) of a base (for example, triethylamine or 37% ammonia, etc.) or acid (such as acetic acid, etc.) are adjusted.

Purified compounds used in prep-TLC, column chromatography or prep-HPLC systems, the solvent system is generally: dichloromethane and methanol, petroleum ether and ethyl acetate, dichloromethane and ethyl acetate, petroleum ether and dichloro For systems such as methane, ethyl acetate and methanol, the volume ratio of the solvent is adjusted depending on the polarity of the compound, and a small amount (0.1% to 10%) of a base (for example, triethylamine or 28% aqueous ammonia, etc.) may be added or The acid (for example, acetic acid, etc.) is adjusted.

Example 1:

Racemic 2-(hydroxymethyl)-2-(methoxymethyl)quinuclidin-3-one

3-quinuclidinone hydrochloride (50.0g), potassium carbonate (50.0g) and 37% formaldehyde (105mL) were mixed in a system of methanol and water (methanol 600mL, water 200mL) at 25 ° C, heated to 70 After reacting for 5 h at ° C, the reaction system was cooled to room temperature, the pH was adjusted to 12-13 with 2N aqueous sodium hydroxide solution, extracted with dichloromethane, and concentrated to give a crude product which was purified by column chromatography to give 2-(hydroxymethyl)-2- (Methoxymethyl)quinuclidin-3-one, 24.0 g, yield 40%.

MS (ESI), m / z , 200.1 [M + H] +.

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm) 3.96 (d, J = 11.8 Hz, 1H), 3.88 - 3.73 (m, 3H), 3.39 (s, 3H), 3.37 - 3.25 (m, 2H), 3.06–2.88 (m, 3H), 2.39 (dd, J=6.0, 3.0 Hz, 1H), 2.10–1.94 (m, 4H).

Example 2:

2-(Hydroxymethyl)-2-(methoxymethyl)quinuclidin-3-one (chiral enantiomer 1)

Chiral enantiomer 1

The racemic 2-(hydroxymethyl)-2-(methoxymethyl)quinuclidin-3-one was separated and separated by chiral column CHIRALPAK AS-H (10mm x 250mm, 5μm), column temperature At 40 ° C, mobile phase A was 0.1% DEA (diethylamine) in Hexane (n-hexane) (v / v), mobile phase B was ethanol, running time was 30 minutes, gradient was mobile phase A / mobile phase B (90 /10,v/v), the flow rate is 6.0 mL/min, the detection wavelength is UV 310 nm, and the separation is collected at a peak RT (retention time) of 12.2 minutes to obtain a single configuration of the compound, which is 2-(hydroxyl). 2-(methoxymethyl)quinuclidin-3-one (enantiomer 1), detected by chiral column CHIRALPAK AS-H (4.6mm x 250mm), column temperature 40 ° C, mobile phase A is 0.1% DEA in Hexane (v/v), mobile phase B is ethanol, running time is 15 minutes, gradient is mobile phase A / mobile phase B (90/10), flow rate is 1.0 mL / min, detection wavelength is UV 310nm, RT 6.1min, ee value 98%.

MS (ESI), m / z , 200.1 [M + H] +.

¹ H-NMR (400 MHz, CDCl ₃ ) δ (ppm) 3.98 (d, J = 11.8 Hz, 1H), 3.88 - 3.75 (m, 3H), 3.41 (s, 3H), 3.33 (ddd, J = 16.6, 7.8, 4.0 Hz, 2H), 3.06 - 2.84 (m, 3H), 2.42 (p, J = 3.0 Hz, 1H), 2.15 - 1.91 (m, 4H).

Example 3:

2-(Hydroxymethyl)-2-(methoxymethyl)quinuclidin-3-one (chiral enantiomer 2)

Chiral enantiomer 2

The racemic 2-(hydroxymethyl)-2-(methoxymethyl)quinuclidin-3-one was separated and separated by chiral column CHIRALPAK AS-H (10mm x 250mm, 5μm), column temperature At 40 ° C, mobile phase A is 0.1% DEA in Hexane (v / v), mobile phase B is ethanol, run time is 30 minutes, gradient is mobile phase A / mobile phase B (90/10, v / v), flow rate The detection wavelength was UV 310 nm at 6.0 mL/min, and the other single configuration was obtained by separation and collection at a peak RT of 14.0 minutes, which was 2-(hydroxymethyl)-2-(methoxymethyl). Quinine cyclo-3-one (enantiomer 2), detected by chiral column CHIRALPAK AS-H (4.6mm x 250mm), column temperature 40 ° C, mobile phase A 0.1% DEA in Hexane (v/v) The mobile phase B was ethanol, the running time was 15 minutes, the gradient was mobile phase A/mobile phase B (90/10), the flow rate was 1.0 mL/min, the detection wavelength was UV 310 nm, RT 7.0 min, and the ee value was 95%.

MS (ESI), m / z , 200.1 [M + H] +.

¹ H-NMR (400 MHz, CDCl ₃ ) δ (ppm) 3.97 (d, J = 11.8 Hz, 1H), 3.88 - 3.74 (m, 3H), 3.44 - 3.36 (m, 3H), 3.38 - 3.27 (m, 2H), 3.05 - 2.82 (m, 3H), 2.40 (dd, J = 6.0, 3.0 Hz, 1H), 2.14 - 1.93 (m, 4H).

Example 4-19:

The compounds of Examples 4 to 19 and their characterization data were prepared according to the synthesis methods of Examples 1 to 3 and the chiral resolution conditions and the corresponding starting materials, and the characterization data thereof were as follows, wherein the chiral enantiomer 1 and the hands were used. The enantiomers 2 are each a pair of enantiomers of the corresponding compound which have different retention times under the chiral resolution conditions of the invention:

Experimental Example 20 H1299 cell proliferation activity test

Cell proliferation activity was tested by the following method:

Materials and instruments used in this experiment:

RPMI1640 (CORNING-CELLGRO, #10-041-CVR)

Fetal Bovine Serum (BIOSERA, #FB-1280)

CellTiter-Glo Luminescent Cell Viability Assay (Promega, #G7572)

96-well plate (Corning, #3788)

96-well plate (Corning, #3797)

96-well plate,black(Corning,#3904)

Backing Tape,white(PE,#6005199)

DMSO (Sigma, #D2650)

Experimental methods and steps

Day 1 H1299 cell plating

1. Trypsinize and determine cell density;

2. Dilute the cell slurry to the desired volume at an optimized density;

3. Dispense 90 μl/well of cell slurry onto the assay plate;

4. Incubate the cells for 24 hours at 37 ° C, 5% CO ₂ and humid conditions.

Day 2 Add compound

1. Prepare the reference compound and test it in 100% DMSO solution according to the plate diagram (final concentration 200x);

2. Transfer 5 μl of the compound to a 95 μl intermediate plate (concentration 10x);

3. Transfer 10 μl of the compound to the test plate (concentration 1x);

4. Incubate at 37 ° C, 5% CO ₂ and humid conditions.

Day 5 Imaging

1. Balance the analytical reagent at room temperature for 30 minutes before use;

2. Add 30 μl of cell titer-Glo reagent to each well and shake the plate for 10 minutes to induce cell lysis;

3. Incubate for 2 minutes to stabilize the illuminating signal;

4. Read the luminescence data on Envision at an interval of 0.5 s.

data processing

Inhibition rate = (Max signal-Compound signal) / (Max signal-Min signal) × 100

Max signal: data from the reading of the solvent DMSO;

Compound signal: data from the action of reading compounds;

Min signal: data from the action of reading the medium;

The cell proliferation activity of the compound of the present invention was measured by the above experimental method, and the compound cell inhibitory activity (IC ₅₀ ) was measured as shown in the following table: + indicates >100 μM, ++ indicates 50-100 μM, and +++ indicates 10-50 μM, + +++ means 1-10 μM and +++++ means <1 μM.

The experimental results show that the inhibitory activity of chiral compounds on tumor cells is significantly better than that of racemic compounds (Example 3 vs. Example 1), and the chiral isomers of different chiral configurations have significant inhibitory activities on tumor cells. Differences (Example 3 vs. Example 2; Example 7 vs. Example 6; Example 11 vs. Example 10; Example 13 vs. Example 12; Example 17 vs. Example 16; Example 19 Relative to Example 18).

Experimental Example 21 Determination of rat pharmacokinetics

Experimental method

SD rats were used as test animals, and the drug concentration in plasma at different times after intravenous administration and intragastric administration of the compound of the example was determined by LC/MS/MS method to study the pharmacokinetics of the compound of the present invention in rats. Learn behavior and evaluate its pharmacokinetic characteristics.

2. Experimental protocol

2.1 Test drugs:

Compounds of the present invention, 1, 2, 3, 10, 11, 14, 15

2.2 Test animals

Healthy adult SD (Sprague-Dawley) rats, male, 7-10 weeks old, weighing 190-260 g, provided by Beijing Weitong Lihua Experimental Animal Co., Ltd., animal certificate number: 11400700226017 and 11400700231083.

2.3 Preparation of test drugs

An appropriate amount of the sample was weighed and dissolved in DMSO/solutol/water = 10/10/80 at a concentration of 2.50 mg/mL.

2.4 Administration of test drugs

Male SD rats each had three test compounds, and were administered overnight after fasting. The intravenous dose was 5 mg/kg and the intragastric dose was 10 mg/kg.

3. Experimental operation

Blood was taken from the jugular vein before and after 0.083-24 h at the time of administration. K2-EDTA was anticoagulated, centrifuged, and plasma was taken and stored at -70 °C until LC/MS/MS analysis.

4. Pharmacokinetic data results

Note: A: IV (5mg/kg); B: PO (10mg/kg).

The experimental results show that the two chiral isomers obtained after resolution show very significant differences in C _max , AUC and F% (bioavailability), and the oral AUC ratio of chiral compounds in one configuration The oral AUC of its enantiomer is more than doubled (Example 3 vs. Example 2; Example 11 vs. Example 10; Example 15 vs. Example 14), one of the configurations of the hand The bioavailability of the sex isomer is 15% or more higher than the bioavailability of its enantiomer (Example 3 vs. Example 2; Example 11 vs. Example 10; Example 15 vs. Example) 14). The chiral compound (the compound of Example 3) was significantly superior to the racemic compound (the compound of Example 1) in terms of _Cmax , AUC, and F% (bioavailability) as compared with the racemic compound.

While the invention has been described with respect to the preferred embodiments of the embodiments of the embodiments of the invention modify. Accordingly, the scope of the invention is defined by the appended claims.

Claims (12)

1. A compound of the formula I-A or I-B, a pharmaceutically acceptable salt, isomer thereof or mixture, solvate, polymorph, stable isotope derivative or prodrug thereof:

   

   among them,

   R is selected from substituted or unsubstituted, unbranched or branched, saturated or unsaturated C _1-10 alkyl or C _3-12 cycloalkyl; substituted or unsubstituted aryl; substituted or unsubstituted a saturated or unsaturated heterocyclic ring; -L ¹ -L ² , L ^{1 is} selected from a substituted or unsubstituted C _1-10 alkyl group, and L ^{2 is} selected from an aryl group or a heterocyclic ring in which the hetero atom is selected from N , O, S or P.
2. A compound of the formula IA or IB, a pharmaceutically acceptable salt, an isomer thereof, or a mixture, solvate, polymorph, stable isotope derivative or prodrug thereof, according to claim 1. It is characterized in that R is selected from a substituted or unsubstituted C _1-10 alkyl group, preferably a substituted or unsubstituted C _1-6 alkyl group.
3. A compound of the formula IA or IB, a pharmaceutically acceptable salt, an isomer thereof, or a mixture, solvate, polymorph, stable isotope derivative or prodrug thereof, according to claim 1. It is characterized in that R is selected from -L ¹ -L ² , L ^{1 is} selected from a substituted or unsubstituted C _1-6 alkyl group, more preferably a C _1-4 alkyl group; and L ² is preferably an aryl group or a heterocyclic ring.
4. A compound of the formula IA or IB, a pharmaceutically acceptable salt, an isomer thereof, or a mixture, solvate, polymorph, stable isotope derivative or prodrug thereof, according to claim 1. Characterized in that R is selected from a methyl group.
5. A compound of the formula IA or IB, a pharmaceutically acceptable salt, an isomer thereof, or a mixture, solvate, polymorph, stable isotope derivative or prodrug thereof, according to claim 1. Characterized in that the compound represented by Formula IA or IB is selected from any of the following structures:

   

   

   

   
6. A compound of the formula IA or IB, a pharmaceutically acceptable salt, an isomer thereof, or a mixture, solvate, polymorph, stable isotope derivative or prodrug thereof, according to claim 1. It is characterized in that the compound represented by the formula I is obtained by separately collecting the compound 1 under chiral resolution conditions at an RT of 12.2 minutes or 14.0 minutes;

   

   Or, the compound of the formula I is obtained by separately collecting the compound 2 under chiral resolution conditions at an RT of 13.2 minutes or 15.4 minutes;

   

   Or, the compound of the formula I is obtained by separately collecting the compound 3 under chiral resolution conditions at an RT of 15.2 minutes or 16.9 minutes;

   

   Or, the compound of formula I is compound 4 collected under chiral resolution conditions at an RT of 14.8 minutes or 17.8 minutes;

   

   Or, the compound of the formula I is obtained by separately collecting the compound 5 under chiral resolution conditions at an RT of 12.5 minutes or 14.4 minutes;

   

   Or, the compound of the formula I is obtained by separately collecting the compound 6 under chiral resolution conditions at an RT of 14.3 minutes or 16.2 minutes;

   

   Or, the compound of the formula I is obtained by separately collecting the compound 7 under chiral resolution conditions at an RT of 12.9 minutes or 14.6 minutes;

   

   Or, the compound of the formula I is obtained by separately collecting the compound 8 under chiral resolution conditions at an RT of 10.5 minutes or 12.1 minutes;

   

   Or, the compound of the formula I is obtained by separately collecting the compound 9 under chiral resolution conditions at an RT of 8.9 minutes or 10.7 minutes;

   

   The chiral splitting conditions include:

   The chiral column is CHIRALPAK AS-H 10mm x 250mm, 5μm;

   The column temperature is 40 ° C;

   Mobile phase A is 0.1% DEA in Hexane, and the percent is volume percent;

   Mobile phase B is ethanol;

   The gradient is mobile phase A / mobile phase B = 90/10, the ratio is the volume ratio;

   The flow rate is 6.0 mL/min;

   The detection wavelength was UV 310 nm.
7. A compound of the formula IA or IB according to any one of claims 1 to 6, a pharmaceutically acceptable salt thereof, an isomer thereof or a mixture thereof, a solvate, a polymorph, and a stable A method for preparing an isotope derivative or prodrug, which comprises the steps of:

   

   The 3-quinuclidinone is reacted with formaldehyde and alcohol ROH under basic conditions to obtain a racemic compound of the formula IC, and the general formula IC can be chiralized under chiral column or chemical resolution. a compound as shown in Formulas IA and IB;

   Wherein the R group is as defined in any one of claims 1-6.
8. A pharmaceutical composition comprising an effective amount of a compound of formula IA or IB, a pharmaceutically acceptable salt, isomer thereof or mixture thereof, according to any one of claims 1-6. Solvates, polymorphs, stable isotope derivatives or prodrugs, and pharmaceutically acceptable carriers, diluents or excipients.
9. A compound of the formula IA or IB, a pharmaceutically acceptable salt, an isomer thereof or a mixture, solvate, polymorph, stable isotope thereof, according to any one of claims 1 to Use of a derivative or prodrug, or a pharmaceutical composition according to claim 8, for the preparation of a medicament for the prevention, alleviation and/or treatment of a p53 mutation-related disease.
10. The use according to claim 9, wherein the p53 mutation-associated disease is selected from the group consisting of cancer, inflammatory disease, autoimmune disease, cardiovascular disease, heart disease or viral infection.
11. The use according to claim 10, wherein said cancer is selected from the group consisting of ovarian cancer, prostate cancer, myeloma, esophageal cancer, gastric cancer, acute myeloid leukemia, chronic myelomonocytic leukemia, acute or chronic leukemia.
12. A compound of the formula IA or IB according to any one of claims 1 to 6, a pharmaceutically acceptable salt thereof, an isomer thereof or a mixture thereof, a solvate, a polymorph, and a stable Use of an isotopic derivative or prodrug, or a pharmaceutical composition according to claim 8 in combination with another anti-cancer agent, preferably an alkylating agent, a platinum drug, a topology Isomerase inhibitors, DNA/RNA synthesis inhibitors, DNA methyltransferases, liposomes, alkaloids, antibody drugs, hormone anticancer agents, proteasome inhibitors, HDAC inhibitors, CDK kinase inhibitors, VEGFR or EGFR inhibitor, m-TOR inhibitor, PI3K kinase inhibitor, B-Raf inhibitor, PARP inhibitor, c-Met kinase inhibitor, ALK kinase inhibitor, AKT inhibitor, ABL inhibitor, FLT3 inhibitor, PD -1 mAb or PD-L1 mAb.