CN109867675B - Pyrrolopyrimidine derivative compound, pharmaceutical composition and application thereof - Google Patents

Abstract

The invention provides a pyrrolopyrimidine derivative compound shown as a general formula (I) or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotopic marker, isomer or prodrug thereof. The invention also provides a pharmaceutical composition containing the same and application of the compound and the pharmaceutical composition in preparing medicines for treating JAK kinase mediated diseases. The pyrrolopyrimidine derivative compound and the pharmaceutical composition thereof provided by the invention have remarkable JAK kinase inhibitory activity, excellent pharmacokinetic properties and low dose, so that the pyrrolopyrimidine derivative compound and the pharmaceutical composition thereof can be administered at a low dose, and can reduce the treatment cost and possible toxic and side effects of patients, thereby having great application potential.

Description

Pyrrolopyrimidine derivative compound, pharmaceutical composition and application thereof

Technical Field

The invention relates to the field of medicines, and particularly relates to a pyrrolopyrimidine derivative compound, a pharmaceutical composition containing the compound and application of the compound as a janus kinase inhibitor.

Background

Janus kinases (JAK) are non-receptor tyrosine kinases (PTK), and the family members have 7 homologous regions (JH 1-7), wherein a JH1 region is a kinase region, and a JH2 region is a pseudo-kinase region. Unlike other PTKs, JAKs lack the Src homology 2(SH2) structure, which can be activated by either catalyzing tyrosine phosphorylation of cytokine receptors linked to them, or by phosphorylating a variety of signaling molecules containing a specific SH2 domain. There are 4 family members of Janus kinases, JAK1, JAK2, TYK2 and JAK3, respectively. The first 3 are widely present in various tissues and cells, while JAK3 is present only in the bone marrow and lymphatic system. The JAK-STAT signal pathway is a cell signal conduction pathway which is discovered in recent years and closely related to inflammatory cytokines and tumors, and is widely involved in important biological processes such as cell proliferation, differentiation, metastasis and apoptosis, immune response regulation, cell homeostasis and the like in human health and disease processes. Cytokine stimulation signals are conducted through the JAK-STAT pathway. The JAK-STAT pathway conducts extracellular chemical signals through the cell membrane to gene promoters located on DNA within the nucleus, ultimately affecting changes in DNA transcription and activity levels in the cell. The JAK-STAT pathway consists of three major components: 1) a receptor; 2) janus kinases (JAK) and 3) Signal Transducer and Activator of Transcription (STAT). The receptor may be activated by interferons, interleukins, growth factors or other chemical messengers, activation leading to autophosphorylation of JAKs; the STAT proteins then bind to phosphorylated receptors, allowing STAT to be phosphorylated by JAKs; phosphorylated STAT proteins are then isolated from the receptor, dimerized, and translocated into the nucleus to bind to specific DNA sites and alter transcription (Scott, m.j., c.j. godshall et al (2002). "Jaks, STATs, Cytokines, and septis" Clin Diagn Lab Immunol 9(6): 1153-9).

The janus kinase family plays a role in cell proliferation and functional cytokine-dependent regulation of immune responses. It is mainly used for screening the therapeutic drugs for rheumatoid arthritis, blood system diseases, tumors, other immunological diseases and the like. At present, many inhibitors of Janus kinases or related kinases have been reported, and Tofacitinib and Ruxolitinib have been approved by FDA for marketing.

Disclosure of Invention

An object of the present invention is to provide a novel compound or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotopic label, isomer or prodrug thereof, which has excellent JAK kinase inhibitory activity.

It is another object of the present invention to provide a pharmaceutical composition.

It is another object of the present invention to provide the use of the novel compounds or pharmaceutically acceptable salts, solvates, active metabolites, polymorphs, isotopic labels, isomers or prodrugs thereof.

The invention provides a pyrrolopyrimidine derivative compound shown as a general formula (I) or pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotopic marker, isomer or prodrug thereof,

wherein the content of the first and second substances,

l is selected from-S (═ O)₂-or-C (═ O) -;

x is selected from CR₄Or N;

R₁selected from the following substituted or unsubstituted groups: halogen, amino, C_1～8Alkyl radical, C_1～8Alkoxy radical, C_2-8Alkenyl radical, C_2-8Alkynyl, C_3～8Cycloalkyl radical, C_3～8Heterocyclic group, C_6～20Aryl or C_5～20A heteroaryl group; the substituent is selected from halogen and C_1～8Alkyl radical, C_1～8Haloalkyl, C_1～8Alkoxy radical, C_3～8Cycloalkyl radical, C_3～8Heterocyclic group, C_6～20Aryl radical, C_5～20Heteroaryl, cyano, amino, hydroxy, carboxy or mercapto;

R₂is selected from-NHR₇Hydroxy or mercapto, wherein R₇Selected from the following substituted or unsubstituted groups: hydrogen, halogen, C_1～6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_1～6Alkyl acyl radical, C_1～6Alkylsulfonyl radical, C_1～6Haloalkyl or C_1～6An alkoxy group; the substituent is selected from cyano, amino, hydroxyl, carboxyl, sulfydryl and C_3～6Cycloalkyl radical, C_3～8Heterocyclic group, C_6～20Aryl or C_5～20A heteroaryl group;

R₃selected from hydrogen, halogen, C_1～6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_1～6Haloalkyl, C_1～6Alkoxy, cyano, amino, hydroxy, carboxyl or mercapto;

R₄selected from hydrogen, halogen or C_1～6An alkyl group;

R₅selected from cyano, -CONH₂Or a carboxyl group;

R₆selected from hydrogen, halogen or C_1～6An alkyl group.

The inventors have studied the JAK kinase inhibitor, barcitinib, and found that: the formula a is the dominant conformation of Baricitinib, and after amino, hydroxyl, sulfydryl and other substituents are introduced to a pyrrole ring, hydrogen atoms on the substituents and nitrogen atoms on pyrimidine can form intramolecular hydrogen bonds (as-NH)₂For example, as shown in formula b), to stabilize this dominant conformation, thereby further improving the biological activity and pharmacokinetic properties of the compound.

In one embodiment according to the present invention, the substituent group R₂Can be selected from-NHR₇Wherein R is₇May be selected from the following substituted or unsubstituted groups: hydrogen, halogen, C_1～4Alkyl radical, C_2-4Alkenyl radical, C_2-4Alkynyl, C_1～4Alkyl acyl radical, C_1～4Alkylsulfonyl radical, C_1～4Haloalkyl or C_1～4An alkoxy group; the substituent is selected from cyano, amino, hydroxyl, carboxyl, mercapto and C_3～5Cycloalkyl radical, C_3～6Heterocyclic group, C_6～12Aryl or C_5～12A heteroaryl group. In a preferred embodiment according to the invention, R₇Can be selected from hydrogen, methyl, ethyl, propyl, isopropyl, n-butyl, methylacyl, ethylaccyl, propylacyl, methylsulfonyl, ethylsulfonyl, propylsulfonyl, methoxy, ethoxy, propoxy, etc., and optional substituents can be fluorine, chlorine, bromine, iodine, cyano, amino, hydroxyl, carboxyl, mercapto, hydroxyl, carboxyl, hydroxyl, or the like,

Phenyl, naphthyl, five-membered or six-membered heterocyclic group or heteroaryl, wherein the heteroatom can be at least one of N, O, S, the number of the substituents can be one or more, and the types can be the same or different.

In one embodiment according to the present invention, the substituent group X may be selected from CH.

In one embodiment according to the present invention, the substituent group R₁May be selected from the following substituted or unsubstituted groups: c_1～6Alkyl radical, C_1～6Alkoxy radical, C_3～6Cycloalkyl radical, C_3～6Heterocyclic group, C_6～12Aryl or C_5～12A heteroaryl group. In a preferred embodiment according to the invention, the substituent R is a substituent₁May be selected from the following substituted or unsubstituted groups: c_1～4Alkyl radical, C_3～5Cycloalkyl, phenyl, biphenyl, naphthyl, C_3～6Heterocyclyl or C_5～6A heteroaryl group. The optional substituents may be halogen or C_1～4The number of the alkyl groups and the substituents may be one or more, and the kinds may be the same or different. In a more preferred embodiment according to the invention, R₁Can be selected from methyl, ethyl, propyl, isopropyl, n-butyl, ethenyl, propenyl, ethynyl, propynyl, methoxy, ethoxy, propoxy,

Phenyl, biphenyl, naphthyl, five-or six-membered heterocyclic or heteroaryl group, wherein the heteroatom may be at least one of N, O, S; optional substituents may be fluoro, chloro, bromo, iodo, methyl, ethyl, propyl, isopropyl, n-butyl.

In one embodiment according to the present invention, the substituent group R₃Can be selected from hydrogen, halogen, C_1～4Alkyl radical, C_2-4Alkenyl radical, C_2-4Alkynyl, C_1～4Haloalkyl, C_1～4Alkoxy, cyano, amino, hydroxyl, carboxyl or mercapto. In a preferred embodiment according to the invention, the substituent R is a substituent₃Can be selected from hydrogen, methyl, ethyl, propyl, isopropyl, n-butyl, cyano, amino, hydroxyl, carboxyl, mercapto, methoxy, ethoxy, propoxy, and trifluoromethyl.

In one embodiment according to the present invention, the substituent group R₅Can be selected from cyano. In one embodiment according to the present invention, the substituent group R₆May be selected from hydrogen. The compounds provided by the present invention are preferably selected from the following structures:

the present invention also provides a pharmaceutical composition comprising a compound of any of the above claims, or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotopic label, isomer, or prodrug thereof, and a pharmaceutically acceptable carrier.

The pharmaceutical composition includes, but is not limited to, oral dosage forms, parenteral dosage forms, external dosage forms, rectal dosage forms, and the like. For example, the pharmaceutical composition may be tablets, capsules, pills, powders, sustained-release preparations, solutions and suspensions for oral administration, sterile solutions, suspensions or emulsions for parenteral injection, ointments, creams, gels, etc. for external use, or suppositories for rectal administration.

The pharmaceutical composition may also include other active ingredients or drugs in combination with the compound or a pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotopic label, isomer or prodrug thereof.

The invention also provides the compound or pharmaceutically acceptable salt, solvate, active metabolite, polymorph, isotopic marker, isomer or prodrug thereof, and application of the pharmaceutical composition in preparing a medicament for treating JAK kinase-mediated diseases. Among them, JAK kinases may include JAK1, JAK2, JAK3 and the like.

Further, the JAK kinase-mediated disease may be a tumor or an autoimmune disease.

Further, diseases mediated by the JAK kinases include, but are not limited to, neoplastic diseases such as leukemia, lymphoma, myeloproliferative neoplasm, myelodysplastic syndrome, and the like; but also autoimmune diseases such as rheumatoid arthritis, psoriatic arthritis, autoimmune skin diseases, and the like.

Further, the JAK kinase-mediated diseases may include, but are not limited to, autoimmune skin diseases such as psoriasis, atopic dermatitis, vitiligo, pruritus, scleroderma, alopecia areata, alopecia totalis, alopecia universalis, androgenetic alopecia, and the like.

In particular, the lymphoma according to the present invention may include, but is not limited to, Hodgkin disease (Hodgkins disease) or Non-Hodgkin lymphoma (Non-Hodgkins leukemia), including, but not limited to, B-cell lymphoma (B-cell lymphoma) or T-cell lymphoma (T-cell lymphoma).

The leukemia of the present invention includes, but is not limited to, Acute lymphocytic leukemia (Acute lymphocytic leukemia), Chronic lymphocytic leukemia (Chronic lymphocytic leukemia), Acute myelogenous leukemia (Acute myelogenous leukemia), and Chronic myelogenous leukemia (Chronic myelocytic leukemia).

Myelodysplastic Syndrome (Myeloproliferative Syndrome) and Myeloproliferative neoplasms (Myeloproliferative neoplasms) described herein include, but are not limited to, Myelofibrosis (Myelofibrosis), Primary Myelofibrosis (Primary Myelofibrosis), Primary thrombocytosis Myelofibrosis (Post-infectious thrombosis Myelofibrosis), Polycythemia Vera Myelofibrosis (Post-Polycythemia Vera Myelofibrosis), Primary thrombocytosis (Essential thrombocytosis), Polycythemia Vera (Polycythemia Vera), Multiple Myeloma (Multiple myelomyelomas).

The tumor provided by the invention comprises solid tumors such as non-small cell lung cancer, liver cancer, kidney cancer, prostate cancer, breast cancer, pancreatic cancer, stomach cancer, colon cancer, melanoma, head and neck cancer and the like besides lymphoma and leukemia.

The autoimmune diseases described herein include, but are not limited to, Rheumatoid Arthritis (Rheumatoid Arthritis), Juvenile Idiopathic Arthritis (Juvenile Idiopathic Arthritis), Juvenile Dermatomyositis (Juvenile Dermatopathy), Atopic Dermatitis (Atopic Dermatitis), Giant Cell Arteritis (Giant Cell Arteritis), Takayasu Arteritis (Takayasu Arteritis), Kawasaki Disease (Kawasaki Disease), Herpes Zoster (Herpes Zoster), Autoinflammatory Disease (Autoinflimatomy synomes), Ankylosing Spondylitis (Ankylating sponalitis), Airesdi-Gouti Syndrome (Aicarvaceous Gouti Syndrome), Alopecia Areata (Alidepicted in the drawings), Alopecia Areata (Alopecia Totalis), Alopecia Areata (Alisma Toxic Toxicosis), Alopecia Areata (Alopecia Areata), Alopecia Areata's, Alopecia Areata (Betulosis), and Alopecia Areata's Disease

Syndrome), CANDLE Syndrome (Chronic inflammatory dermatological Dermatology With Lipodystrophy and elongated Temperation), Graft-versus-host Disease (Graft-cover-host Disease), Diabetic renal Disease (Diabetic Kidney Disease), Dry Eye Syndrome/Keratoconjectionosis Sicca, Non-infectious Uveitis (Non-infectious Uveitis), Pruritis (Pruritis), psoriasis (Psori)(iii) asis), Plaque Psoriasis (Plaque Psoriasis), Psoriatic Arthritis (Psoriatic Arthritis), Inflammatory Bowel Disease (Inflammatory Bowel Disease), Crohn's Disease (Crohn's Disease), Small-intestine Crohn's Disease (Small Boweh Crohn's Disease), fistula Crohn's Disease (fistuling Crohn's Disease), Ulcerative Colitis (Ulctive Colitis), SVAI Disease (Stimulator of Interferon Genes (STING) -Associated Vasculopathy With Ulcerative Colitis Infancy), Systemic Lupus Erythematosus (Systemic Lus Erythematosis), Scleroderma (Systemic Sclerosis/Scleroderma), Vitiligo (Vitiligo), and Chronic Beryllium Disease (Chronic Berry Disease).

The pyrrolopyrimidine derivative compound and the pharmaceutical composition thereof provided by the invention have remarkable JAK kinase inhibitory activity, excellent pharmacokinetic properties and low dose, so that the pyrrolopyrimidine derivative compound and the pharmaceutical composition thereof can be administered at a low dose, and can reduce the treatment cost and possible toxic and side effects of patients, thereby having great application potential.

Detailed description of the invention

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. All patents, patent applications, and publications cited herein are incorporated by reference in their entirety unless otherwise indicated. When a trade name appears herein, it is intended to refer to its corresponding commodity or its active ingredient.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the subject matter herein. In this application, it must be noted that, unless the context clearly dictates otherwise, as used in this specification and the claims, the singular forms "a," "an," and "the" include plural referents. It should also be noted that the use of "or", "or" means "and/or" unless stated otherwise. Furthermore, the term "comprising" as well as other forms, such as "includes," "including," and "containing," are used without limitation.

Definitions of standardized chemical terms can be found in literature work, including "Advanced Organic Chemistry 4" by Carey and Sundberg^th Ed,Vol A(2000)and B(2001),Plenum Press, New York. Unless otherwise indicated, conventional methods within the skill of the art are employed, such as mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques, and pharmacological methods. Unless a specific definition is set forth, the nomenclature and laboratory procedures and techniques related to analytical chemistry, synthetic organic chemistry, and chemistry such as medical and pharmaceutical chemistry are known to those skilled in the art. Standard techniques can be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, drug delivery and treatment of patients. Standard techniques can be used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipid infection). For example, the reaction and purification techniques can be carried out using a kit with instructions provided by the manufacturer, or according to methods known in the art, or according to the methods described herein. In general, the foregoing techniques and procedures may be practiced by conventional methods well known in the art and described in various general or more specific documents that are cited and discussed in this disclosure.

When a substituent is described by a general formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the formula is written from right to left. For example, CH₂O is equivalent to OCH₂。

The terms "optionally" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not. For example, ethyl is "optionally" substituted with halo, meaning that ethyl may be unsubstituted (-CH)₂CH₃) Monosubstituted (e.g. -CH)₂CH₂F) Polysubstituted (e.g. -CHFCH)₂F、-CH₂CHF₂Etc.) or fully substituted (-CF)₂CF₃). It will be appreciated by those skilled in the art that any group containing one or more substituents will not incorporate any substitution or substitution pattern which is sterically impossible and/or cannot be synthesized.

The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced with a substituent, so long as the valence of the particular atom is normal and the substituted compound is stable. When the substituent is oxo (i.e., ═ O), meaning that two hydrogen atoms are substituted, oxo does not occur on the aryl.

When any variable (e.g., R) occurs more than one time in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2R, the group may optionally be substituted with up to two R, and there are separate options for R in each case. Furthermore, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

As used herein C_m～nMeaning that the moiety has m to n carbon atoms. For example, the "C" is_1～8By "group" is meant a moiety having from 1 to 8 carbon atoms, i.e., the group contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms … … 8 carbon atoms. Thus, for example, "C_1～8Alkyl "means an alkyl group containing 1 to 8 carbon atoms, i.e., the alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl … … octyl, and the like. Herein, numerical ranges, such as "1 to 8" refer to each integer in the given range, such as "1 to 8 carbon atoms" means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, or 8 carbon atoms.

The term "alkyl" refers to an optionally substituted straight or optionally substituted branched chain saturated aliphatic hydrocarbon group attached to the rest of the molecule by a single bond. The "alkyl" groups herein may have from 1 to about 8 carbon atoms, for example from 1 to 6 carbon atoms, or from 1 to 4 carbon atoms or from 1 to 3 carbon atoms. Examples of "alkyl" herein include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, 2-methyl-l-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-l-butyl, 2-methyl-3-butyl, 2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-l-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl4-methyl-2-pentyl, 2-dimethyl-l-butyl, 3-dimethyl-1-butyl, 2-ethyl-1-butyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, hexyl and the like, as well as longer alkyl groups such as heptyl and octyl and the like. When a group as defined herein, such as "alkyl" appears in a numerical range, e.g. "C_1-8Alkyl "refers to an alkyl group that can be composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, and as a further example," C_1-4Alkyl "refers to an alkyl group that can be made up of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms. Alkyl herein also includes the case where no numerical range is specified.

The term "alkenyl" refers to an optionally substituted straight or optionally substituted branched monovalent hydrocarbon radical having at least one C ═ C double bond. The alkenyl group has, but is not limited to, 2 to about 8 carbon atoms, such as 2 to about 6 carbon atoms, 2 to about 4 carbon atoms. The double bond in these groups may be in either the cis or trans conformation and should be understood to encompass both isomers. Examples of alkenyl groups include, but are not limited to, ethenyl (CH ═ CH)₂) 1-propenyl (CH)₂CH＝CH₂) Isopropenyl (C (CH)₃)＝CH₂) Butenyl, 1, 3-butadienyl and the like. When a numerical range is present for alkenyl as defined herein, e.g. "C_2-8The "alkenyl group" means an alkenyl group which may be composed of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, and the alkenyl group herein also covers the case where a numerical range is not specified.

The term "alkynyl" refers to an optionally substituted, straight or branched chain, monovalent hydrocarbon radical having at least one C ≡ C triple bond. The alkynyl group has, but is not limited to, 2 to about 8 carbon atoms, such as 2 to about 6 carbon atoms, or 2 to about 4 carbon atoms. Examples of alkynyl groups herein include, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, and 1, 3-butadiynyl, and the like. When a numerical range occurs for alkynyl as defined herein, for example "C_2-8Alkynyl means a group consisting of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atomsCarbon atom, an alkynyl group of 8 carbon atoms, alkynyl herein also encompasses instances where no numerical range is specified.

The term "cycloalkyl" refers to a non-aromatic carbon-containing ring, including saturated carbocycles (e.g., cycloalkyl) or unsaturated carbocycles (e.g., cycloalkenyl). Carbocycle includes monocyclic (having one ring), and may be, for example, monocyclic cycloalkyl; a bicyclic carbocycle (having two rings), for example, may be a bicyclic cycloalkyl; carbocyclic (having more than two rings). The rings may be bridged or spiro. Carbocycle (e.g., cycloalkyl or cycloalkenyl) can have 3 to 8 carbon atoms, for example, 3 to about 6 ring-forming carbon atoms or 3 to about 5 ring-forming carbon atoms.

The term "aryl" refers to optionally substituted aromatic hydrocarbon groups having from 6 to about 20, such as 6 to 12 or 6 to 10 ring-forming carbon atoms, which may be monocyclic aryl, bicyclic aryl or higher ring aryl. The bicyclic aryl or higher ring aryl may be a monocyclic aryl fused to other independent rings such as alicyclic, heterocyclic, aromatic ring, aromatic heterocyclic. Non-limiting examples of monocyclic aryl groups include monocyclic aryl groups of 6 to about 12, 6 to about 10, or 6 to about 8 ring-forming carbon atoms, such as phenyl; bicyclic aryl is for example naphthyl; polycyclic aryl radicals are, for example, phenanthryl, anthracyl, azulenyl.

The term "heteroaryl" refers to optionally substituted heteroaryl groups containing from about 5 to about 20, such as 5 to 12 or 5 to 10, backbone ring-forming atoms, wherein at least one (e.g., 1-4, 1-3, 1-2) ring-forming atoms is a heteroatom independently selected from the group consisting of heteroatoms of oxygen, nitrogen, sulfur, phosphorus, silicon, selenium and tin, but is not limited thereto. Heteroaryl includes monocyclic heteroaryl (having one ring), bicyclic heteroaryl (having two rings), or polycyclic heteroaryl (having more than two rings). In embodiments where two or more heteroatoms are present in the ring, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may be different from each other. The bicyclic heteroaryl or higher ring heteroaryl may be a monocyclic heteroaryl fused with other independent rings such as alicyclic ring, heterocyclic ring, aromatic heterocyclic ring (which may be collectively referred to as fused ring heteroaryl). Non-limiting examples of heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolinyl, isoquinolinyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothienyl, indolyl, isoindolyl, and the like.

The term "heterocyclyl" refers to a non-aromatic heterocyclic ring, which includes saturated or unsaturated heterocyclic rings (containing unsaturation), does not have a completely conjugated pi-electron system, and can be classified as a monocyclic, fused polycyclic, bridged or spiro ring system without aromaticity. Wherein one or more (e.g., 1-4, 1-3, 1-2) ring-forming atoms are heteroatoms, such as oxygen, nitrogen or sulfur atoms. Heterocycles can include mono-heterocycles (having one ring) or bis-heterocycles (having two bridged rings) or polyheterocycles (having more than two bridged rings); spiro rings are also included. A heterocyclyl group can have 3 to about 20 ring-forming atoms, such as 3 to about 10, 3 to about 8, 4 to 7, 5 to about 8, or 5 to about 6 ring-forming atoms. Non-limiting examples of heterocyclyl groups include oxiranyl, thietanyl, aziridinyl, azetidinyl, oxetanyl, thietanyl, tetrahydrofuryl, pyrrolidinyl, oxazolidinyl, tetrahydropyrazolyl, pyrrolinyl, dihydrofuranyl, dihydrothienyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyranyl, dihydrothiopyranyl, azepanyl, oxepanyl, thiepanyl, thiepinyl, oxaazabicyclo [2.2.1] heptyl, and azaspiro [3.3] heptyl groups and the like.

The term "halo" or "halogen" refers to an optionally substituted group (e.g., alkyl, alkenyl, alkynyl, alkoxy, etc.) wherein at least one hydrogen atom is replaced with a halogen (e.g., fluorine, chlorine, bromine, iodine, or combinations thereof). In some embodiments, two or more hydrogens are replaced with the same halogen as each other (e.g., difluoromethyl, trifluoromethyl); in other embodiments two or more hydrogens are replaced with halogens that are not exactly the same as each other (e.g., 1-chloro-1-fluoro-1-iodoethyl).

The term "alkoxy" refers to an alkyl ether group (O-alkyl), non-limiting examples of which include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy and the like.

The term "alkanoyl" refers to a group wherein an alkyl group is attached to the group-CO-, non-limiting examples of which include formyl, acetyl, propionyl, butyryl and the like. For example, the term "C_1-6Alkylacyl "means C_1-6Alkyl groups are linked to-CO-to form groups. As another example, the term "C_1-4Alkylacyl "means C_1-4Alkyl groups are linked to-CO-to form groups.

The term "alkylsulfonyl" refers to alkyl and-SO₂-linked to form a group, non-limiting examples of which include methylsulfonyl, ethylsulfonyl, propylsulfonyl, butylsulfonyl and the like. For example, the term "C_1-6Alkylsulfonyl "means C_1-6Alkyl and-SO₂-linked to form a group. As another example, the term "C_1-4Alkylsulfonyl "means C_1-4Alkyl and-SO₂-linked to form a group.

The term "amino" refers to the group-NH₂Group, -NH (C)_1～6Alkyl) group or-N (C)_1～6Alkyl radical)₂A group. Specific examples of amino groups include, but are not limited to, -NH₂、-NHCH₃、-N(CH₃)₂、-NHC₂H₅、-N(C₂H₅)₂、-N(C₃H₇)₂、-N(CH₃)C₂H₅And the like.

Other group terms herein also include: "hydroxy" refers to the group-OH, "mercapto" refers to the group-SH, "cyano" refers to the group-CN, and "carboxy" refers to the group-COOH.

The term "member" refers to the number of backbone atoms that make up a ring. For example, pyridine is a six-membered ring and pyrrole is a five-membered ring.

The term "pharmaceutically acceptable" is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutical composition" refers to a biologically active compound optionally mixed with at least one pharmaceutically acceptable chemical ingredient or agent, i.e., a "carrier," which aids in the introduction of the compound into cells or tissues, including but not limited to stabilizers, diluents, suspending agents, thickening agents, and/or excipients.

The term "pharmaceutically acceptable salt" refers to salts that retain the biological potency of the free acid and free base of the specified compound, and that are biologically or otherwise not adversely affected. As the salt in the present invention, metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like can be mentioned unless otherwise specified. Non-limiting examples of metal salts include, but are not limited to, salts of alkali metals, such as sodium, potassium, and the like; salts of alkaline earth metals such as calcium, magnesium, barium, and the like; aluminum salts, and the like. Non-limiting examples of salts with organic bases include, but are not limited to, salts with trimethylamine, triethylamine, pyridine, picoline, 2, 6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, and the like. Non-limiting examples of salts with inorganic acids include, but are not limited to, salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like. Non-limiting examples of salts with organic acids include, but are not limited to, salts with formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, malic acid, maleic acid, tartaric acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like. Non-limiting examples of salts with basic amino acids include, but are not limited to, salts with arginine, lysine, ornithine, and the like. Non-limiting examples of salts with acidic amino acids include, but are not limited to, salts with aspartic acid, glutamic acid, and the like.

Pharmaceutically acceptable salts can be synthesized from the parent compound, which contains an acid or base, by conventional chemical methods. In general, such salts are prepared by the following method: prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid, in water or an organic solvent or a mixture of the two. Generally, nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

The term "solvate" refers to a physical aggregate of a compound of the present invention with one or more solvent molecules, the physical aggregate including varying degrees of ionic and covalent bonding, such as hydrogen bonding. It has been shown that this solvate can be isolated, for example, when one or more solvent molecules are incorporated into the crystal lattice of the crystal. "solvate" includes both a solvent phase and an isolatable solvate moiety. Examples of corresponding solvates are numerous and include ethanol solvates, methanol solvates, and the like. "hydrate" is a compound of water (H)₂O) the molecule is a solvate of the solvent. One or more of the compounds of the present invention may be optionally prepared as solvates. The preparation of solvates is well known. For example, M.Caira et al, J.pharmaceutical Sci, 93(3),601-611(2004) describe the preparation of solvates of the antifungal agent fluconazole, i.e., using ethyl acetate and water. Van binder et al, AAPS pharmcitech, 5(1), article 12 (2004); similar preparation of solvates, hydrates are also described in a.l. bingham et al, chem.commu., 603-604 (2001). A typical, non-limiting, preparation procedure is to dissolve the compound of the invention in the desired amount of the desired solvent (organic solvent or water or a mixture thereof) at a temperature above ambient temperature, cool it down, settle it for crystallization, and then isolate and pick out the crystals by standard methods. The presence of the solvent (water) forming the solvate (hydrate) in the crystallization can be confirmed by i.r. spectroscopic analysis technique.

The term "active metabolite" refers to an active derivative of a compound that is formed upon metabolism of the compound.

The term "polymorph" refers to a compound of the invention in different crystal lattice forms.

The term "isotopic label" refers to isotopically labeled compounds of the present invention. For example, isotopes in the compounds of the invention may include various isotopes of H, C, N, O, P, F, S, etc., such as²H,³H,¹³C,¹⁴C,¹⁵N,¹⁸O,¹⁷O,³¹P,³²P,³⁵S,¹⁸F and³⁶S。

the term "pharmaceutically acceptable prodrug" or "prodrug" refers to any pharmaceutically acceptable salt, ester, salt of an ester, or other derivative of a compound of the invention that, upon administration to a recipient, is capable of providing, directly or indirectly, a compound of the invention or a pharmaceutically active metabolite or residue thereof. Particularly preferred derivatives or prodrugs are those compounds which, when administered to a patient, enhance the bioavailability of the compounds of the present application (e.g., may allow an orally administered compound to be more readily absorbed into the blood), or facilitate delivery of the parent compound to a biological organ or site of action (e.g., the brain or lymphatic system). Prodrugs can be prepared by modifying functional groups present in the compound in a manner that results in the modification of the parent compound by conventional procedures or in vivo. Various prodrug forms are well known in the art. See, Pro-drugs as Novel Delivery Systems (1987) Vol.14of the A.C.S.Sympossium Series, Bioreproducible Carriers in Drug Design, (1987) Edward B.Roche, ed., American Pharmaceutical Association, and Pergamon Press, both by T.Higuchi and V.Stella. Design of produgs, Bundgaard,

ed, Elseview,1985and Method in Enzymology, Widder, k.et al, ed; academy, 1985, vol.42, p.309-396; bundgaard, H. "Design and Application of drugs" in A Textbook of Drug Design and Development, Krosgaard-Larsen and H. Bundgaard, Ed.,1991, Chapter 113 and 191; and Bundgaard, h., Advanced Drug Delivery Review,1992,8,1-38, which are incorporated herein by reference.

The term "stereoisomer" refers to an isomer resulting from the different arrangement of atoms in a molecule. The compounds of the present invention contain asymmetric or chiral centers, double bonds, etc., and thus, the compounds of the present invention may include various isomer forms such as optical isomers, geometric isomers, tautomers, atropisomers, etc., and these isomers and single isomers, racemates, etc. thereof are included in the scope of the present invention. For example, for optical isomers, optically active (R) -and (S) -isomers, and D and L isomers, can be prepared by chiral resolution, chiral synthesis, or chiral reagents, or other conventional techniques. For example, they may be converted to diastereomers by reaction with a suitable optically active substance (e.g., a chiral alcohol or Mosher's moh's acid chloride), separated and converted (e.g., hydrolyzed) to the corresponding single isomers. For another example, the separation may be performed by a chromatography column.

The "pharmaceutical compositions" herein may be prepared in a manner well known in the pharmaceutical arts and may be administered or administered by a variety of routes depending on whether local or systemic treatment is desired and the area to be treated. Can be administered topically (e.g., transdermal, dermal, ocular, and mucosal including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal), orally, or parenterally. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intracerebroventricular, administration. The administration may be parenteral in a single bolus form, or may be by, for example, a continuous infusion pump. The pharmaceutical compositions herein include, but are not limited to, the following forms: tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (solid or dissolved in a liquid vehicle); ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions, sterile packaged powders and the like containing, for example, up to 10% by weight of the active compound.

The pharmaceutical compositions herein may be formulated in unit dosage forms, each dosage form containing from about 0.1 to about 1000mg, typically from about 5 to about 1000mg, more typically from about 100 to about 500mg of active ingredient. The term "unit dosage form" refers to physically discrete single dosage units suitable as unitary dosages for human patients and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical carrier.

The term "subject" refers to a subject, including mammals and non-mammals, suffering from a disease, disorder or condition, and the like. Examples of mammals include, but are not limited to, any member of the class mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs, and the like.

The term "treating" and other similar synonyms include alleviating, alleviating or ameliorating a symptom of a disease or disorder, preventing other symptoms, ameliorating or preventing the underlying metabolic cause of the symptom, inhibiting the disease or disorder, e.g., arresting the development of the disease or disorder, alleviating the disease or disorder, ameliorating the disease or disorder, alleviating a symptom caused by the disease or disorder, or halting a symptom of the disease or disorder, and further, the term can be included for prophylactic purposes. The term also includes obtaining a therapeutic effect and/or a prophylactic effect. The therapeutic effect refers to curing or ameliorating the underlying disease being treated. In addition, a cure or amelioration of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect, e.g., an improvement in the condition of the patient is observed, although the patient may still be affected by the underlying disease. For prophylactic effect, the composition or compound can be administered to a patient at risk of developing a particular disease, or to a patient presenting with one or more physiological symptoms of the disease, even if a diagnosis of the disease has not yet been made.

The term "a dose necessary to achieve a therapeutic effect" or "therapeutically effective amount" refers to an amount of at least one agent or compound sufficient to alleviate to some extent one or more symptoms of the disease or condition being treated after administration. The result may be a reduction and/or alleviation of signs, symptoms, or causes, or any other desired change in a biological system. An effective amount suitable in any individual case can be determined using techniques such as a dose escalation assay. The amount of the compound, pharmaceutical composition or medicament actually administered will generally be determined by a physician, in the light of the relevant circumstances, including the condition being treated, the chosen route of administration, the actual compound administered; age, weight and response of the individual patient; severity of patient symptoms, etc.

The proportion or concentration of the compound of the invention in the pharmaceutical composition may not be fixed and will depend on a variety of factors including dosage, chemical properties (e.g., hydrophobicity), route of administration, and the like. For example, the compounds of the present invention can be provided for parenteral administration by a physiological buffered aqueous solution containing about 0.1-10% w/v of the compound. Some typical dosage ranges are from about 1. mu.g/kg to about 1g/kg body weight/day. In certain embodiments, the dosage range is from about 0.01mg/kg to about 100mg/kg body weight/day. The dosage will likely depend on such variables as the type and extent of progression of the disease or disorder, the general health status of the particular patient, the relative biological efficacy of the selected compound, the excipient formulation and its route of administration.

The term "administering" refers to a method capable of delivering a compound or composition to a desired site for biological action. These methods include, but are not limited to, oral routes, via the duodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical and rectal administration. Administration techniques useful for The compounds and methods described herein are well known to those skilled in The art, for example, in Goodman and Gilman, The pharmaceutical Basis of Therapeutics, current ed.; pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.

The term "IC₅₀By "is meant that 50% inhibition of the maximal effect is obtained in the assay measuring such effect.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, technical solutions of exemplary embodiments of the present invention will be further described below.

The compounds of the present invention can be prepared by the following methods. The following methods and examples are intended to illustrate these methods. These schemes and examples should not be construed as limiting the invention in any way. The compounds described herein can also be synthesized using standard synthetic techniques known to those skilled in the art, or using methods known in the art in combination with those described herein.

The chemical reactions of the embodiments of the present invention are carried out in a suitable solvent that is compatible with the chemical changes of the present invention and the reagents and materials required therefor. In order to obtain the compounds of the present invention, it is sometimes necessary for a person skilled in the art to modify or select the synthesis steps or reaction schemes based on the existing embodiments.

An important consideration in any synthetic route planning in the art is the selection of suitable protecting groups for reactive functional groups, such as amino groups in the present invention. Greene and Wuts (Protective Groups In Organic Synthesis, Wiley and Sons,1991) are the authorities of this area for trained practitioners. All references cited herein are incorporated herein in their entirety.

The reactions described herein can be monitored according to any suitable method known in the art. For example, it can be determined by a broad spectrum method such as nuclear magnetic resonance spectroscopy (e.g.¹H or¹³C) Infrared spectroscopy, spectrophotometry (e.g., UV-visible light), mass spectrometry, etc., or monitoring product formation by chromatography, such as High Performance Liquid Chromatography (HPLC) or thin layer chromatography.

The partial compounds of the general formula I according to the invention can be prepared by those skilled in the art of organic synthesis by the following procedures using standard methods in the art:

wherein, compound 1 and compound 2 are subjected to Michael addition reaction to generate compound 3, and compound 3 is subjected to SEM protecting group removal on amino to obtain compound I of the invention.

Example 12- (3- (3-amino-4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (benzene Formyl) azetidin-3-yl) acetonitrile

Step A: 2- ((1-benzoyl) azetidin-3-ylidene) acetonitrile

300mg (2.3mmol,1.0eq) of 2- (azetidin-3-ylidene) acetonitrile hydrochloride and 700mg (6.9mmol,3.0eq) of triethylamine were dissolved in 20mL of dichloromethane, and 387mg (2.7mmol,1.2eq) of benzoyl chloride was slowly added dropwise to the above reaction solution at room temperature. The reaction was allowed to stand overnight at room temperature. The reaction was quenched with 30mL of saturated aqueous sodium bicarbonate, the aqueous phase was extracted with dichloromethane, the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The residue was purified by column chromatography (PE: EA: 2:1 to 1:1) to give the objective compound (397mg, yield: 87%).

And B: 4-chloro-7- ([2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidine

38.4g (250.4mmol,1.0eq.) of 4-chloro-7H-pyrrolo [2,3-d ] pyrimidine are dissolved in 200mL of dry DMF solution with stirring in an ice bath, and 13g (305mmol,1.2eq) of NaH with a 57% content are added. The reaction was stirred at room temperature for 1 hour, and 50.9g of SEMCl (305mmol,1.2eq.) were added dropwise with cooling in an ice bath. After the addition was completed, the reaction was stirred in ice bath for 1 hour, quenched with water, extracted with ethyl acetate, and the combined organic phases were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo, and subjected to column chromatography on silica gel to obtain the objective compound (71g, yield ═ 100%).

And C: 2-cyano-2- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) acetic acid ethyl ester

33.5g (118mmol,1.0eq.) of 4-chloro-7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidine are added, with stirring at room temperature, to a mixture of 40.1g (354.0mmol,3.0eq.) of ethyl cyanoacetate and 33.0g (238mmol,2.0eq.) of potassium carbonate. The temperature of the reactants is raised to 60 ℃ for reaction for 0.5 hour, and then the temperature is raised to 130 ℃ for reaction for 1.0 hour. Cooling to room temperature, quenching with water, extraction with ethyl acetate, washing the combined organic phases with brine, drying over sodium sulfate, filtering and concentrating in vacuo, and column chromatography on silica gel afforded the title compound (30.6g, yield 72%).

Step D: 2- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) acetonitrile

50g (855.6mmol,10.0eq.) of sodium chloride was added to a mixed solvent of 30.6g (84.9mmol,1.0eq.) of ethyl 2-cyano-2- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) acetate in DMSO and water with stirring at room temperature. Reacting at 150 ℃ for 6 days under the protection of liquid nitrogen and gas. Cooling to room temperature, quenching with water, extraction with ethyl acetate, washing the combined organic phases with brine, drying over sodium sulfate, filtering and concentrating in vacuo, and column chromatography on silica gel afforded the title compound (18.1g, 74% yield).

Step E: 4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -3-amino-1H-pyrazole

37g of hydrazine hydrate (628mmol,10.0eq.) with a content of 85% were added to 18.1g (62.8mmol,1.0eq.) of 2- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrole [2,3-d ] at room temperature with stirring]Pyrimidin-4-yl) acetonitrile and 22.4g (188mmol,3eq.) of DMF-DMA in 80mL of DMF. The reaction was stirred and refluxed at 90 ℃ for 3 hours under the protection of liquid nitrogen. After cooling to room temperature, 100mL of water was added, followed by stirring, suction filtration, and drying, the objective compound (11.0g, yield: 53%) was obtained.¹HNMR(400MHz,DMSO-d₆)δ12.13(brs,1H),8.65(s,1H),8.18(brs,1H),7.61(d,J＝3.2Hz,1H),7.01(d,J＝3.6Hz,1H),6.55(brs,2H),5.60(s,2H),3.52(t,J＝8.0Hz,2H),0.83(t,J＝8.0Hz,2H),-0.10(s,9H).LC-MS:m/z＝331[M+1]⁺。

Step F: 2- (3- (3-amino-4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (benzoyl) azetidin-3-yl) acetonitrile

335mg (1.0mmol,1.0eq) of 4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-3-amine and 200mg (1.0mmol,1.0eq) of 2- ((1-benzoyl) azetidin-3-ylidene) acetonitrile were dissolved in 10mL of acetonitrile, and 310mg (2.0mmol,2.0eq) of 1, 8-diazabicyclo [5.4.0] undec-7-ene were slowly added dropwise to the reaction system at room temperature, and the reaction was stirred at room temperature overnight. The reaction was quenched with 30mL of a saturated aqueous sodium bicarbonate solution, the aqueous phase was extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and the residue was subjected to column chromatography (PE: EA ═ 2:1 to EA) to give the target compound (405mg, yield ═ 76%).

Step G: 2- (3- (3-amino-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (benzoyl) azetidin-3-yl) acetonitrile

405mg (1.0mmol,1.0eq) of 2- (3- (3-amino-4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] are introduced at room temperature]Pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (benzoyl) azetidin-3-yl) acetonitrile was dissolved in a mixture of 8mL dichloromethane and 1mL trifluoroacetic acid. The reaction was carried out at room temperature overnight. The solvent was evaporated under reduced pressure, the residue was dissolved in 10mL of methanol, 0.5mL of ethylenediamine was added at room temperature, and the reaction was stirred at room temperature for 20 minutes. The reaction mixture was poured into 50mL of water, the aqueous phase was extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and the residue was subjected to column chromatography (DCM: EA ═ 1:1 to EA) to give the target compound (120mg, yield ═ 39%).¹H-NMR(400MHz,DMSO-d6):δ12.04(s,1H),8.64(d,J＝12.9Hz,2H),7.71(d,J＝7.5Hz,2H),7.59-7.46(m,4H),7.10(s,1H),6.38(s,2H),4.98(d,J＝9.3Hz,1H),4.65(d,J＝10.4Hz,1H),4.57(d,J＝8.7Hz,1H),4.35(d,J＝10.1Hz,1H),3.63(s,2H).LC-MS:m/z＝399[M+H]⁺.

Example 22- (3- (3-amino-4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (p-pyrimidine Tosyl) azetidin-3-yl) acetonitrile

Step A: 2- (1- (p-toluenesulfonyl) azetidin-3-ylidene) acetonitrile

The compound was prepared with reference to step a described in example 1 (495mg, yield 87%).

And B: 2- (3- (3-amino-4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (p-toluenesulfonyl) azetidin-3-yl) acetonitrile

Compound (243mg, yield 52%) was prepared with reference to step F described in example 1.

And C: 2- (3- (3-amino-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (p-toluenesulfonyl) azetidin-3-yl) acetonitrile

Compound (70mg, yield: 37%) was prepared with reference to step G described in example 1.¹H-NMR(400MHz,DMSO-d6):δ12.05(s,1H),8.64(s,1H),8.25(s,1H),7.71(d,J＝7.5Hz,2H),7.58(s,1H),7.32(d,J＝7.7Hz,2H),7.01(s,1H),6.25(s,2H),4.31(d,J＝9.7Hz,2H),4.13(d,J＝9.8Hz,2H),3.42(s,2H),2.03(s,3H).LC-MS:m/z＝449[M+H]⁺.

Example 32- (3- (3-amino-4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (ortho-pyrazole Propylsulfonyl) azetidin-3-yl) acetonitrile

Step A: 2- (1- (n-propylsulfonyl) azetidin-3-ylidene) acetonitrile

Compound (383mg, yield 76%) was prepared with reference to step a described in example 1.

And B: 2- (3- (3-amino-4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (n-propylsulfonyl) azetidin-3-yl) acetonitrile

Compound (374mg, yield 72%) was prepared with reference to step F described in example 1.

And C: 2- (3- (3-amino-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (n-propylsulfonyl) azetidin-3-yl) acetonitrile

Compound (140mg, yield ═ 50%) was prepared with reference to step G described in example 1.¹H-NMR(400MHz,DMSO-d6):δ12.06(s,1H),8.67(s,1H),8.62(s,1H),7.56(s,1H),7.10(s,1H),6.39(s,2H),4.55(d,J＝8.9Hz,2H),4.14(d,J＝8.8Hz,2H),3.58(s,2H),3.23-3.16(m,2H),1.79-1.68(m,2H),1.00(t,J＝7.3Hz,3H).LC-MS:m/z＝401[M+H]⁺.

Example 42- (3- (3-amino-4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (cyclo Propylsulfonyl) azetidin-3-yl) acetonitrile

Step A: 2- (1- (cyclopropylsulfonyl) azetidin-3-ylidene) acetonitrile

Compound (396mg, yield 79%) was prepared with reference to step a described in example 1.

And B: 2- (3- (3-amino-4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (cyclopropylsulfonyl) azetidin-3-yl) acetonitrile

Compound (368mg, yield 69%) was prepared with reference to step F described in example 1.

And C: 2- (3- (3-amino-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (cyclopropylsulfonyl) azetidin-3-yl) acetonitrile

A compound was prepared with reference to step G described in example 1 (165mg, yield 59%).¹H-NMR(400MHz,DMSO-d6):δ12.06(s,1H),8.67(s,1H),8.63(s,1H),7.56(s,1H),7.11(s,1H),6.40(s,2H),4.62(d,J＝9.1Hz,2H),4.18(d,J＝9.0Hz,2H),3.60(s,2H),2.90-2.78(m,1H),1.09-0.97(m,4H).LC-MS:m/z＝399[M+H]⁺.

Example 52- (3- (3-amino-4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (ethyl) Sulfonyl) azetidin-3-yl) acetonitrile

Step A: 2- (1- (ethylsulfonyl) azetidin-3-ylidene) acetonitrile

The compound is prepared by referring to the patent US2009/233903A1Page 63-64.

And B: 2- (3- (3-amino-4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (ethanesulfonyl) azetidin-3-yl) acetonitrile

Compound (1.05g, yield 85%) was prepared with reference to step F described in example 1.

And C: 2- (3- (3-amino-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (ethanesulfonyl) azetidin-3-yl) acetonitrile

The compound was prepared with reference to step G described in example 1 (384mg, yield ═ 49%).¹H-NMR(400MHz,DMSO-d6):δ12.06(s,1H),8.67(s,1H),8.61(s,1H),7.56(s,1H),7.10(s,1H),6.39(s,2H),4.56(d,J＝8.9Hz,2H),4.13(d,J＝9.0Hz,2H),3.59(s,2H),3.25-3.16(m,2H),1.25(t,J＝7.1Hz,3H).LC-MS:m/z＝387[M+H]⁺.

Example 62- (3- (3-amino-4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (iso-forms Propylsulfonyl) azetidin-3-yl) acetonitrile

Step A: 2- (1- (isopropylsulfonyl) azetidin-3-ylidene) acetonitrile

Compound (506mg, yield 100%) was prepared with reference to step a described in example 1.

And B: 2- (3- (3-amino-4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (isopropylsulfonyl) azetidin-3-yl) acetonitrile

Compound (316mg, yield 60%) was prepared with reference to step F described in example 1.

And C: 2- (3- (3-amino-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (isopropylsulfonyl) azetidin-3-yl) acetonitrile

Compound (100mg, yield 43%) was prepared with reference to step G described in example 1.¹H-NMR(400MHz,DMSO-d6):δ12.05(s,1H),8.67(s,1H),8.61(s,1H),7.55(s,1H),7.09(s,1H),6.40(s,2H),4.56(d,J＝8.7Hz,2H),4.10(d,J＝8.7Hz,2H),3.60(s,2H),3.38-3.33(m,1H),1.28(s,3H),1.26(s,3H).LC-MS:m/z＝401[M+H]⁺.

Example 72- (3- (3-amino-4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (ortho-pyrazole Butylsulfonyl) azetidin-3-yl) acetonitrile

Step A: 2- (1- (n-butylsulfonyl) azetidin-3-ylidene) acetonitrile

A compound (517mg, yield 95%) was prepared with reference to step a described in example 1.

And B: 2- (3- (3-amino-4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (n-butylsulfonyl) azetidin-3-yl) acetonitrile

A compound (321mg, yield 59%) was prepared with reference to step F described in example 1.

And C: 2- (3- (3-amino-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (n-butylsulfonyl) azetidin-3-yl) acetonitrile

Compound (108mg, yield ═ 45%) was prepared with reference to step G described in example 1.¹H-NMR(400MHz,DMSO-d6):δ12.06(s,1H),8.67(s,1H),8.62(s,1H),7.56(s,1H),7.10(s,1H),6.39(s,2H),4.56(d,J＝9.0Hz,2H),4.14(d,J＝9.0Hz,2H),3.58(s,2H),3.25-3.19(m,2H),1.71-1.65(m,2H),1.48-1.34(m,2H),0.89(t,J＝7.2Hz,3H).LC-MS:m/z＝415[M+H]⁺.

Example 82- (1-acetyl-3- (3-amino-4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) -1H-pyrazole-1- Yl) azetidin-3-yl) acetonitrile

Step A: 2- (1- (acetyl) azetidin-3-ylidene) acetonitrile

Compound (327mg, yield 95%) was prepared with reference to step a described in example 1.

And B: 2- (3- (3-amino-4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (acetyl) azetidin-3-yl) acetonitrile

Compound (328mg, yield 70%) was prepared with reference to step F described in example 1.

And C: 2- (3- (3-amino-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (acetyl) azetidin-3-yl) acetonitrile

Compound (36mg, yield ═ 16%) was prepared with reference to step G described in example 1.¹H-NMR(400MHz,DMSO-d6):δ8.84(s,1H),8.73(s,1H),7.84(d,J＝2.0Hz,1H),7.30(d,J＝2.8Hz,1H),6.50(s,2H),4.83(d,J＝9.3Hz,1H),4.56(d,J＝10.3Hz,1H),4.49(d,J＝9.4Hz,1H),4.21(d,J＝10.3Hz,1H),3.70(s,2H),1.94(s,3H).LC-MS:m/z＝337[M+H]⁺.

Example 92- (3- (3, 5-diamino-4- (7H-pyrrolo [2,3-d ]]Pyrimidin-4-yl) -1H-pyri dineAzol-1-yl) - 1- (ethylsulfonyl) azetidin-3-yl) acetonitrile

Step A: 2- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) malononitrile

10.0g (35.2mmol,1.0eq.) of 4-chloro-7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidine are added, with stirring at room temperature, to a mixture of 3.5g (53mmol,1.5eq.) of malononitrile and 7.2g (53mmol,1.5eq.) of potassium carbonate. The reaction was warmed to 60 ℃ for 6 hours. Cooling to room temperature, quenching with water, extraction with ethyl acetate, washing the combined organic phases with brine, drying over sodium sulfate, filtering and concentrating in vacuo, and column chromatography on silica gel afforded the title compound (9.3g, 84% yield).

And B: 4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -3, 5-diamino-1H-pyrazole

7.5g of hydrazine hydrate (127mmol,10.0eq.) with a content of 85% are added to 4.0g (12.8mmol,1.0eq.) of 2- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrole [2,3-d ] with stirring at room temperature]Pyrimidin-4-yl) malononitrile in 100mL of absolute ethanol. The reaction was stirred under reflux at 90 ℃ overnight under the protection of liquid nitrogen. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure, filtered with suction, and dried to obtain the objective compound (1.98g, yield ═ 45%). LC-MS M/z 346[ M + 1]]⁺。

And C: 2- (3- (3, 5-diamino-4- (7- ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (ethanesulfonyl) azetidin-3-yl) acetonitrile

Compound (130mg, yield 75%) was prepared with reference to step F described in example 1.

Step D: 2- (3- (3, 5-diamino-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -1H-pyrazol-1-yl) -1- (ethanesulfonyl) azetidin-3-yl) acetonitrile

Compound (54mg, yield 61%) was prepared with reference to step G described in example 1. LC-MS M/z 402[ M + H ]]⁺.

Examples 10 to 16

The following compounds were synthesized in a similar manner to the above examples.

Example 172- (3- (3-amino-4- (9H-purin-6-yl) -1H-pyrazol-1-yl) -1- (ethylsulfonyl) azepine Cyclobut-3-yl) acetonitrile

4-chloro-7H-pyrrolo [2,3-d ] is prepared by following the procedures described in example 1 and example 5]Pyrimidine was replaced with 6-chloropurine to prepare the title compound (64 mg). LC-MS M/z 388[ M + H ]]⁺.

Examples 18 to 24

The following compounds were synthesized in a similar manner to the above examples.

Example 252- (3- (3, 5-diamino-4- (9H-purin-6-yl) -1H-pyrazol-1-yl) -1- (ethanesulfonyl) Azetidin-3-yl) acetonitrile

4-chloro-7H-pyrrolo [2,3-d ] by reference to the procedures described in example 1 and example 9]Pyrimidine was replaced with 6-chloropurine to prepare the title compound (73 mg). LC-MS, M/z 403[ M + H ]]⁺.

Examples 26 to 32

The following compounds were synthesized in a similar manner to the above examples.

Evaluation of effects

₅₀1. Enzymatic activity (IC) assay

The activity of the compound is determined by establishing a JAK1/2/3 kinase activity detection platform by adopting Lance Ultra principle, and the known Baricitinib is determined to be used as a control. In the assay plate, the enzyme, Ulight-labeled polypeptide substrate, ATP and assay compound are mixed and the reaction is incubated. After the reaction, EDTA is added to stop the reaction, and Eu-labeled antibody is added at the same time for detection. The assay plate was analyzed using Envision from PE in TR-FRET format and data are expressed as readings of 665nm and 615nm, respectively, fluorescence signals. Wherein a high ratio of 665nm/615nm indicates a high enzyme activity, and a low ratio of 665nm/615nm indicates a low enzyme activity.

Reagent: kinase (JAK1/2/3), substrate (ULight-JAK-1peptide and ATP), detection reagent (Eu-W1024Anti-phosphotyrosine and EDTA).

The instrument comprises the following steps: echo, Envision.

The test compounds were dissolved in 10mM DMSO and stored in a nitrogen cabinet for a long period of time. 10 μ L of 10mM test compound solution was diluted to 1mM working solution and diluted 3-fold with Echo for a total of 11 concentrations. The concentration of the compound in the final reaction system was 10. mu.M to 0.17 nM. Adding 5 mu L of enzyme and polypeptide substrate mixed solution into a detection plate by using an electric pipettor, centrifuging the detection plate, and incubating the detection plate at room temperature (23 ℃) for 15 minutes; mu.L of kinase buffer containing ATP was added to the assay plate using an electric pipette, the assay plate was centrifuged, and the assay plate was incubated at room temperature (23 ℃ C.) for 90 minutes using an aluminum foil seal plate. Stopping the reaction, adding a detection reagent into a detection plate by using an electric pipettor, centrifuging the detection plate, and placing the detection plate at room temperature (23 ℃) for half an hour by using an aluminum foil sealing plate; the signal value of the reaction plate was measured by Envision instrument. The results are shown in Table 1.

TABLE 1 inhibitory Activity of Compounds on JAK/1/2/3

The results in Table 1 show that the compound of the invention has good inhibitory activity on JAK/1/2/3 enzyme and is obviously superior to the existing compound Baricitinib.

2.Pharmacokinetic experiments

Female SD rats were grouped into 3 groups, and positive compound Baricitinib and test compound sample suspensions (10mg/kg) were administered orally in a single gavage, or positive compound Baricitinib (1mg/kg) and test compound sample supernatant (2mg/kg) were administered intravenously, respectively. Animals were fasted overnight prior to the experiment, with the fasting time ranging from 10 hours prior to dosing to 4 hours post-dosing. Blood was collected at 0.25, 0.5, 1, 2, 4, 8, 12 and 24 hours after oral administration in the oral group, and at 0.083, 0.25, 0.5, 1, 2, 4, 8 and 24 hours after oral administration in the intravenous group. After isoflurane anesthesia by using a small animal anesthesia machine, 0.3mL of whole blood is collected through an eyeground venous plexus, the whole blood is placed in a heparin anticoagulation tube, a sample is centrifuged at 4 ℃ and 4000rpm for 5min, and plasma is transferred to a centrifuge tube and stored at-80 ℃ until analysis. Samples from plasma were extracted using protein precipitation and the extracts were analyzed by LC/MS/MS. The results of the pharmacokinetic experiments are shown in tables 2 and 3.

TABLE 2 pharmacokinetic parameters after gastric gavage of rats with 10mg/kg of different compounds

TABLE 3 pharmacokinetic parameters after intravenous administration of different Compounds in rats

As can be seen from the results in tables 2 and 3, the pharmacokinetic properties of the compound of example 6 of the present invention are also significantly better than that of the existing compound, Baricitinib. The pharmacokinetic performance of the compounds of the remaining examples of the invention, tested in the same manner, was also significantly better than that of Baricitinib.

3. Determination of efficacy in a mouse subcutaneous xenograft tumor model

SPF grade CB-17SCID female mice, 4-5 weeks old, were divided into groups of 5 mice per group. 0.1ml of HEL cell suspension (containing 5.0X 10) suspended in serum-free medium⁶cells, 30% Matrigel) were injected subcutaneously into the left and right underarm area of each mouse. Average tumor volume of more than 100mm³When the method is used, the mice are marked according to the serial numbers, the tumor sizes and the body weights of the mice are respectively measured, the mice are randomly grouped according to the tumor volumes from small to large, and the average body weights of the animals in each group are also adjusted properly to be at the same level. Oral administration was started on the same day in groups, during which tumor volume and body weight were measured 2 times per week, using tumor growth inhibition rate (TGI) as an index of detection.

The tumor growth inhibition rate was calculated as TGI (%) ═ 1- (tumor volume in treatment group-tumor volume in treatment group)/(tumor volume in control group-tumor volume in control group)) × 100%

The results are shown in Table 4.

TABLE 4 in vivo efficacy model in mice

As can be seen from the results in Table 4, the compound of example 6 of the present invention has a good tumor-inhibiting effect on mice, and is significantly superior to the existing compound, Baricitinib, at the same dosage. The compounds of the other examples of the invention were tested in the same manner and the resulting tumor suppression was also significantly better than that of Baricitinib.

4. Treating atopic dermatitis (eczema)

The compound of example 6 is used as an active ingredient to prepare an ethanol solution with the content of 1%, and the ethanol solution is smeared on an affected part of eczema once a day, and the affected part is observed to be obviously improved on the 8 th day and the 20 th day after the medicine is taken.

Although the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims.

Claims (12)

1. A pyrrolopyrimidine derivative compound represented by general formula (I) or a pharmaceutically acceptable salt thereof,

wherein the content of the first and second substances,

l is selected from-S (═ O)₂-or-C (═ O) -;

x is selected from CR₄Or N;

R₁selected from the following substituted or unsubstituted groups: c_1～8Alkyl radical, C_1～8Alkoxy radical, C_2-8Alkenyl radical, C_2-8Alkynyl, C_3～8Cycloalkyl radicals、C_3～8Heterocyclic group, C_6～20Aryl or C_5～20A heteroaryl group; the substituent is selected from halogen and C_1～8Alkyl radical, C_1～8Haloalkyl, C_1～8Alkoxy radical, C_3～8Cycloalkyl radical, C_3～8Heterocyclic group, C_6～20Aryl radical, C_5～20Heteroaryl, cyano, amino, hydroxy, carboxy or mercapto;

R₂is selected from-NHR₇Wherein R is₇Selected from hydrogen;

R₃selected from hydrogen or amino;

R₄selected from hydrogen, halogen or C_1～6An alkyl group;

R₅selected from cyano, -CONH₂Or a carboxyl group;

R₆selected from hydrogen, halogen or C_1～6An alkyl group.

2. A compound or pharmaceutically acceptable salt thereof according to claim 1, wherein X is selected from CH.

3. A compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R is₁Selected from the following substituted or unsubstituted groups: c_1～6Alkyl radical, C_1～6Alkoxy radical, C_3～6Cycloalkyl radical, C_3～6Heterocyclic group, C_6～12Aryl or C_5～12A heteroaryl group; the substituents being selected from halogen or C_1～4An alkyl group.

4. A compound or pharmaceutically acceptable salt thereof according to claim 2, wherein R₁Selected from the following substituted or unsubstituted groups: c_1～6Alkyl radical, C_1～6Alkoxy radical, C_3～6Cycloalkyl radical, C_3～6Heterocyclic group, C_6～12Aryl or C_5～12A heteroaryl group; the substituents being selected from halogen or C_1～4An alkyl group.

5. The compound of any one of claims 1-4 or a pharmaceutically thereofAn acceptable salt, wherein, R₁Is selected from C_1～4Alkyl radical, C_3～5Cycloalkyl, phenyl, biphenyl, naphthyl, C_3～6Heterocyclyl or C_5～6A heteroaryl group.

6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the structures:

7. a pharmaceutical composition comprising a compound of any one of claims 1-6, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

8. Use of a compound of any one of claims 1-6, or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition of claim 7 in the manufacture of a medicament for treating a janus kinase-mediated disease.

9. The use of claim 8, wherein the janus kinase-mediated disease is a tumor or an autoimmune disease.

10. The use of claim 8, wherein the janus kinase-mediated disease is leukemia, lymphoma, myeloproliferative neoplasm, or myelodysplastic syndrome.

11. The use according to claim 8, wherein the janus kinase-mediated disease is rheumatoid arthritis, psoriatic arthritis or autoimmune skin disease.

12. The use according to claim 11, wherein the autoimmune skin disease is psoriasis, atopic dermatitis, vitiligo, pruritus, scleroderma, alopecia areata, alopecia totalis, alopecia universalis, or androgenetic alopecia.