WO2017124934A1 - Phosphine-containing carboline derivative serving as bromodomain inhibitor - Google Patents

Abstract

Provided are a phosphine-containing carboline derivative serving as a bromodomain inhibitor , and a method and drugs for controlling diseases intervened by a bromodomain protein.

Description

Phosphide-containing porphyrin derivative as a domain inhibitor of the bromo region Technical field

The present invention relates to a phosphine-containing porphyrin derivative as a bromodomain inhibitor.

Background technique

The genome of eukaryotes is highly organized within the nucleus. The long chain of double-stranded DNA is entangled with a group of protein octamers (most commonly comprising two copies of histones H2A, H2B, H3 and H4) to form nucleosomes. The basic unit is then further compressed by nucleosome aggregation and folding to form a highly agglomerated chromatin structure. A range of different condensed states are possible, and the tightness of the structure changes during the cell cycle and is most tight during the cell division process. Chromatin structure plays a key role in regulating gene transcription, and gene transcription cannot be efficiently produced by highly condensed chromatin. The chromatin structure is controlled by a series of post-translational modifications to histones, particularly histones H3 and H4, and the modifications are most commonly within the histone tail, which extends beyond the core nucleosome structure. These modifications include acetylation, methylation, phosphorylation, ubiquitination, and SUMOylation. These epigenetic marks are written and erased by specific enzymes that will be tagged to specific residues within the tail of the histone, thus forming an epigenetic code, which is then interpreted by the cell to allow for the chromatin structure. Specific regulation and thus transcription.

Histone acetylation is most commonly associated with activation of gene transcription, as this modification relaxes the interaction of DNA and histone octamers by altering electrostatic properties. In addition to this physical change, specific proteins bind to acetylated lysine residues within histones to read epigenetic coding. In the case of histones, the bromodomains are small, distinct domains within a protein that are normally, but not exclusively, bound to acetylated lysine residues. There are about 50 species of proteins known to contain the Bromo domain domain, and they have a range of functions within the cell.

The Bet family domain-containing protein of the Bet family includes four proteins (BRD2, BRD3, BRD4 and BRD-t) containing a tandem Bromo domain domain, and the tandem Bromo domain domain can bind two The acetylated lysine residue is in close proximity, thereby increasing the specificity of the interaction. It has been reported that BRD2 and BRD3 bind to histones along actively transcribed genes and may be involved in promoting transcriptional elongation (Leroy et al., Mol. Cel. 2008 30(1): 51-60), while BRD4 appears to be involved in pTEF-I3 The complex recruits an inducible gene, resulting in phosphorylation of RNA polymerase and increased transcriptional output (Hargreaves et al Cell, 2009 138(1): 129-145). Moreover, all family members have certain functions in the control or execution of the cell cycle and have been shown to remain complexed with chromosomes during cell division, suggesting a role in maintaining epigenetic memory. In addition, as part of the viral replication process, some viruses utilize these proteins to tether their genome to the chromatin of host cells (You et C. Cell, 2004 117(3): 349-60).

Related recent articles include Trends in Molecular Medicines about BET (2014, 20(9) 477-478); Trends in Pharmacological Sciences (2012, 33(3) 146-153); J. Med. Chem., (2012, 55). ,9393-9413);J.Biol.Chem.,(2 012,287(46):38956). Hundreds of different genetic factors are identified, many of which are proteins associated with chromatin. These associated proteins are directly linked to different diseases such as cancer, neurological diseases, metabolic diseases, cardiovascular diseases, viruses, inflammation, autoimmune diseases and the like. Among the clinically developed Bromo domain domain inhibitors are BMS-986158 (BMS), MRK-8628 (Merck), BAY 1238097 (Bayer), INCB54329 (Incyte) and the like. Thus, the Bromo domain domain inhibitors provided herein provide a means of controlling lesions that are intervened by a Bromo domain domain protein.

Summary of the invention

It is an object of the present invention to provide a novel phosphine-containing porphyrin derivative which is a Bromo zone domain inhibitor.

According to a first aspect of the present invention, there is provided a phosphine-containing porphyrin derivative, a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer thereof, wherein the phosphine-containing porphyrin-derived The structure of the object is as shown in the following formula I:

In the formula,

R ₁ , R ₂ , R ₃ and R ₄ are each independently hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _{1 -8} alkoxy, C _1-8 alkoxycarbonyl, C _3-8 epoxyalkyl, aryl, heteroaryl, or 3- to 12-membered heterocyclic; or, adjacent R ₁ , R ₂ , R ₃ , R _{4 together} with the carbon atom to which they are bonded form a 3- to 9-membered ring;

R _{5 is} selected from the group consisting of hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-8 alkoxy, C _1-8 An alkoxycarbonyl group, an aryl group, a heteroaryl group, or a 3- to 12-membered heterocyclic group;

R _{6 is} selected from the group consisting of hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-8 alkoxy, C _1-8 An alkoxycarbonyl group, an aryl group, a heteroaryl group, or a 3- to 12-membered heterocyclic group;

R _{7 is} selected from the group consisting of: hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-8 alkoxy, C _1-8 An alkoxycarbonyl group, an aryl group, a heteroaryl group, or a 3- to 12-membered heterocyclic group;

R _{8 is} selected from the group consisting of: hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-8 alkoxy, C _1-8 Alkoxycarbonyl;

R _{9 is} selected from the group consisting of: hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-8 alkoxy, C _1-8 Alkoxycarbonyl;

Wherein each of the above alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, 3- to 9-membered rings is optionally and independently substituted by one or more substituents Substituted, each of the substituents is independently hydrazine, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3- to 12-membered Heterocyclyl, aryl, heteroaryl, CN, NO ₂ , =O, =S.

In another preferred embodiment, each substitution refers to substitution with from 1 to 6 substituents, preferably from 1 to 3 substituents.

In another preferred embodiment, the 3- to 9-membered ring optionally contains 1-2 additional heteroatoms selected from N, O or S.

In another preferred embodiment, the 3- to 9-membered ring is saturated or unsaturated.

In another preferred embodiment, R ₁ and R _{2 together} with the carbon atom to which they are attached form a 3- to 9-membered ring, preferably a 5-membered ring or a 6-membered ring.

In another preferred embodiment, R ₂ and R _{3 together} with the carbon atom to which they are attached form a 3- to 9-membered ring, preferably a 5-membered ring or a 6-membered ring.

In another preferred embodiment, R _{4 is} selected from the group consisting of hydrogen, halogen, C _1-3 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _1-5 alkoxy.

In another preferred embodiment, R _{5 is} selected from the group consisting of C _3-8 epoxyalkyl, aryl, and heteroaryl.

In another preferred embodiment, R _{6 is} selected from the group consisting of aryl, heteroaryl, and 3- to 12-membered heterocyclic.

In another preferred embodiment, the 3- to 12-membered heterocyclic group is saturated or unsaturated and contains 1, 2, or 3 heteroatoms selected from N, O or S.

In another preferred embodiment, R _{7 is} selected from the group consisting of hydrogen, halogen, C _1-3 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _1-5 straight chain and branched alkoxy base.

In another preferred embodiment, R _{8 is} selected from the group consisting of hydrogen, halogen, deuterated C _1-3 alkyl, C _2-4 alkenyl, C _2-4 alkynyl.

In another preferred embodiment, R ₈ is a deuterated methyl group.

In another preferred embodiment, R _{9 is} selected from the group consisting of hydrogen, halogen, C _1-8 alkyl, C _1-8 haloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 Cycloalkyl, C _1-8 alkoxy, C _1-8 alkoxycarbonyl.

In another preferred embodiment, R ₉ is a deuterated methyl group.

In another preferred embodiment, R _{9 is} selected from the group consisting of C _1-3 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-5 alkoxy .

In another preferred embodiment, the configuration of the chiral carbon atom in the compound of formula (I) is R or S.

In another preferred embodiment, the phosphine-containing porphyrin derivative is:

According to a second aspect of the present invention, a pharmaceutical composition comprising: (i) an effective amount of the compound of claim 1, or a pharmaceutically acceptable salt thereof; and (ii) A pharmaceutically acceptable carrier.

A third aspect of the invention provides the use of a compound of formula I, or a pharmaceutically acceptable salt thereof, according to the first aspect of the invention, for use in:

(a) preparing a Bromo domain domain inhibitor;

(b) preparing a medicament for preventing and/or treating a disease associated with the Bromo domain; and/or

(c) Non-therapeutic inhibition of the Bromo region domain in vitro.

In another preferred embodiment, the disease is selected from the group consisting of cancer, neurological diseases, metabolic diseases, cardiovascular diseases, viral infections, inflammation, tissue fibrosis-related diseases, and autoimmune diseases.

In another preferred embodiment, the tumor is selected from the group consisting of: lung cancer, breast cancer, blood cancer, cervical cancer, ovarian cancer, intestinal cancer, pancreatic cancer, prostate cancer, liver cancer, brain tumor, skin cancer, and other solid tumors.

In a fourth aspect of the invention, there is provided a method of inhibiting the activity of a Bromo domain, comprising the steps An inhibitory effective amount of a compound of formula I according to the first aspect of the invention, or a pharmaceutically acceptable salt thereof, or an inhibitory effective amount of a pharmaceutical composition according to the second aspect of the invention, administered to an inhibitory subject, .

According to a fifth aspect of the invention, there is provided a process for the preparation of a compound according to the first aspect of the invention, which comprises the steps of:

(1) reacting a compound of formula II with a compound of formula III to form a compound of formula I, wherein L is a leaving group and M is a group that can be coupled to L.

In another preferred embodiment, the method further includes the steps of:

(2) reacting a compound of formula IV with a compound of formula V to form a compound of formula II wherein L is a leaving group.

In another preferred embodiment, the method further includes the steps of:

(3) The step of preparing the formula IV by the compound of the formula VI.

In another preferred embodiment, the method further includes the steps of:

(4) reacting a compound of formula VIII with a compound of formula VII to form a compound of formula VI wherein L is a leaving group and M is a group which is conjugated to L.

In another preferred embodiment, the method further includes the steps of:

(5) A step of preparing a compound of the formula VIII by a compound of the formula IX, wherein Hal is a halogen and M is as defined above.

In another preferred embodiment, the reactivity of Hal ₁ in the step (5) is superior to that of Hal ₂ .

In another preferred embodiment, L is a halogen group.

In another preferred embodiment, M is a boric acid or borate group.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

detailed description

The inventors of the present application have extensively and intensively studied for the first time to develop a novel phosphine-containing porphyrin derivative having the structure shown in Formula I for use as a Bromo domain domain inhibitor. On the basis of this, the present invention has been completed.

the term

Unless otherwise stated, "or" as used herein has the same meaning as "and/or" (refers to "or" and "and").

Unless otherwise specified, among all compounds of the present invention, each chiral carbon atom (chiral center) may optionally be in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.

As used herein, the term "alkyl", alone or as part of another substituent, refers to a straight (ie, unbranched) or branched alkyl group having from 1 to 8 carbon atoms, or a combination thereof. The alkyl group can be saturated, monounsaturated or polyunsaturated, and can include divalent or multivalent radicals. When the alkyl group has a carbon number limitation (for example, C _1-10 ), it means that the alkyl group has 1 to 10 carbon atoms, and for example, the C _1-8 alkyl group may include 1 to 8 carbon atoms. A linear or branched alkyl group such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like.

As used herein, the term "alkenyl", when used alone or as part of another substituent, refers to a straight or branched, carbon chain having at least one carbon-carbon double bond. The alkenyl group having one double bond may be represented by -C _n H _2n-1 , and the alkenyl group having 2 double bonds may be represented by -C _n H _2n-3 . When the alkenyl group has a carbon number limitation (for example, C _2-8 ), it means that the alkenyl group has 2 to 8 carbon atoms, for example, a linear or branched alkenyl group having 2 to 8 carbon atoms. For example, vinyl, propenyl, 1,2-butenyl, 2,3-butenyl, butadienyl, or the like.

As used herein, the term "alkynyl", when used alone or as part of another substituent, refers to an aliphatic hydrocarbon group having at least one carbon to carbon triple bond. The alkynyl group can be straight or branched, or a combination thereof. In some embodiments, the alkynyl group has 2-12 (eg, 2-8, 2-6, or 2-4) carbon atoms. When the alkynyl group has a carbon number number (for example, C _2-8 alkynyl group), it means that the alkynyl group has 2 to 8 carbon atoms. For example, the term "C _2-8 alkynyl group" means having 2 to 8 A linear or branched alkynyl group of one carbon atom, such as ethynyl, propynyl, isopropynyl, butynyl, isobutynyl, sec-butynyl, tert-butynyl, or the like.

As used herein, when used alone or as part of another substituent, the term "cycloalkyl" refers to a monocyclic, bicyclic or tricyclic (including cyclo, bridged or spiro) ring system having a saturated or partially saturated. The cycloalkyl group may have 3 to 12 (e.g., 3 to 10, or 5 to 10) carbon atoms. When a certain cycloalkyl group has a carbon number limitation (e.g., C _3-10 ), it means that the cycloalkyl group has 3 to 10 carbon atoms. In some preferred embodiments, the term "C _3-8 cycloalkyl" refers to a saturated or partially saturated monocyclic or bicyclic alkyl group having from 3 to 8 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentane. A group, a cycloheptyl group, or the like.

As used herein, the term "alkoxy" or "alkyloxy" refers to an alkyl group (eg, -O-alkyl) attached through an oxygen atom, wherein alkyl is as defined above. Examples of specific alkoxy groups are, for example but not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, Or a similar group. The alkoxy group may be substituted by one or more substituents such as a halogen, an amino group, a cyano group, or a hydroxyl group. The alkoxy group can be straight or branched. When the alkoxy group has a carbon number limitation (e.g., C _1-8 ), it means that the cycloalkyl group has 1 to 8 carbon atoms.

As used herein, the term "halogen", when used alone or as part of another substituent, refers to F, Cl, Br, and I.

As used herein, the term "alkoxycarbonyl" refers to a straight or branched alkyl-oxycarbonyl moiety (alkoxy-C=O). The alkoxy group may have 1 to 8 carbon atoms. When the alkoxycarbonyl group has a carbon number limitation (for example, C _1-8 ), it means that the alkyl moiety of the alkoxycarbonyl group has 1-8 carbon atoms, for example, a C _1-8 alkoxycarbonyl group. Refers to a group having a C _1-8 alkoxy-C=O- structure, such as a methoxycarbonyl group, an ethoxycarbonyl group, a tert-butoxycarbonyl group, or the like.

As used herein, the term "aryl", when used alone or as part of another substituent, refers to a monocyclic, bicyclic or fused ring aromatic hydrocarbon group. The aryl group may be substituted or unsubstituted. When an aryl group has a carbon number limit (e.g., C _6-12 ), it means that the aryl group has 6 to 12 carbon atoms. Examples of aryl groups are, for example but not limited to, phenyl, biphenyl, naphthyl, or the like (each of which may be optionally substituted). In the present invention, the aryl group is preferably a C _6-12 aryl group.

As used herein, the term "heteroaryl", when used alone or as part of another substituent, refers to a monocyclic, bicyclic or fused ring aromatic group having a particular number of ring carbon atoms (eg, C _4-10 has 4 to 10 ring-forming carbon atoms) and includes at least one same or different hetero atom selected from N, O or S. The atoms on each ring can be arbitrarily substituted. The heteroaryl group may be 5- to 15-membered, having 1 to 5 aromatic ring groups each independently selected from a hetero atom of N, O or S. Examples of heteroaryl groups are, for example but not limited to, pyridine, pyrimidine, pyrrole, oxazole, indole, furan, benzofuran, thiophene, or the like.

As used herein, when used alone or as part of another substituent, the term "heterocyclyl" refers to a saturated or partially saturated substituent of a monocyclic or fused ring having a particular number of ring-forming carbon atoms (eg, C). _3-11 has 3 to 11 ring-forming carbon atoms) and includes at least one same or different hetero atom selected from N, O or S. In the present invention, the heterocyclic group may be a 3- to 15-membered heterocyclic group having 1 to 5 hetero atoms each independently selected from N, O or S. Examples of the heterocyclic group are, for example but not limited to, a nitrogen heterocyclic group, an oxaheterocyclic group, a thioheterocyclic group, a nitrogen oxyheterocyclyl group, a nitrogen thioheterocyclic group, an oxathioheterocyclic group, etc., more preferably The heterocyclic groups appearing in the various examples of the present application. In the present invention, the heterocyclic group may be monocyclic, bicyclic or tricyclic (including a bicyclic ring, a bridged ring or a spiro ring).

The term "optional" or "optional" (eg, "optionally substituted") as used herein refers to The moiety is substituted or unsubstituted and the substitution only occurs with a chemically achievable position. For example, H, a covalent bond or a -C(=O)- group may not be substituted by a substituent.

As used herein, "oxygen" or "oxy" refers to =0.

As used herein, unless otherwise specified, the term "pharmaceutically acceptable salt" refers to a salt that is suitable for contact with the tissue of a subject (eg, a human) without causing unpleasant side effects. In some embodiments, a pharmaceutically acceptable salt of a compound of the invention includes a salt (eg, a potassium salt, a sodium salt, a magnesium salt, a calcium salt) of a compound of the invention having an acidic group or is basic A salt of a compound of the invention (e.g., a sulfate, a hydrochloride, a phosphate, a nitrate, a carbonate).

As used herein, the term "substituted" (when with or without "optionally") means that one or more hydrogen atoms on a particular group are replaced by a particular substituent. Particular substituents are the substituents described above in the corresponding paragraphs, or the substituents which appear in the examples. Unless otherwise stated, an optionally substituted group may have a substituent selected from a particular group at any substitutable position of the group, and the substituents may be the same or different at each position. A cyclic substituent, such as a heterocycloalkyl group, may be attached to another ring, such as a cycloalkyl group, to form a spirobicyclic ring system, for example, two rings having a common carbon atom. Those skilled in the art will appreciate that the combinations of substituents contemplated by the present invention are those that are stable or chemically achievable. The substituents are, for example but not limited to, hydrazine, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3- to 12-membered Cyclo, aryl, heteroaryl, halogen, hydroxy, carboxy (-COOH), C _1-8 aldehyde, C _2-10 acyl, C _2-10 ester, amino. In the present invention, the deuterated alkyl group may be fully deuterated, or partially deuterated or a mixture thereof.

For convenience and in accordance with conventional understanding, the term "optionally substituted" or "optionally substituted" applies only to sites which are capable of being substituted by a substituent, and does not include those which are not chemically achievable.

Pharmaceutically acceptable salts, solvates, stereoisomers, tautomers

The term "pharmaceutically acceptable salt" as used herein refers to a salt of a compound of the invention and a pharmaceutically acceptable inorganic and organic acid, wherein preferred inorganic acids include, but are not limited to, hydrochloric acid, hydrogen Bromic acid, phosphoric acid, nitric acid, sulfuric acid; preferred organic acids include, but are not limited to: formic acid, acetic acid, propionic acid, succinic acid, naphthalene disulfonic acid (1, 5), asiamic acid, oxalic acid, tartaric acid, lactic acid , salicylic acid, benzoic acid, valeric acid, diethyl acetic acid, malonic acid, succinic acid, fumaric acid, pimelic acid, adipic acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, Gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methanesulfonic acid, p-toluenesulfonic acid, citric acid, and amino acids.

The term "pharmaceutically acceptable solvate" as used herein refers to a compound of the invention that forms a solvate with a pharmaceutically acceptable solvent, wherein the pharmaceutically acceptable solvent includes, but is not limited to, water , ethanol, methanol, isopropanol, tetrahydrofuran, dichloromethane.

The term "pharmaceutically acceptable stereoisomer" as used herein means that the chiral carbon atom to which the compound of the invention relates may be in the R configuration, in the S configuration, or a combination thereof.

Pharmaceutical composition

The invention also provides a pharmaceutical composition having significant anti-tumor efficacy comprising a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable salts Carrier.

Mixing the compound itself or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable excipient, diluent, and the like The preparation is administered orally in the form of a tablet, a capsule, a granule, a powder or a syrup, or is administered orally in the form of an injection. The pharmaceutical composition preferably contains, as an active ingredient, a compound of the formula I of the present invention or a pharmaceutically acceptable salt thereof in an amount of from 0.01% to 99% by weight, more preferably from 0.1% to 90% by weight of the active ingredient.

The above formulations can be prepared by conventional pharmaceutical methods. Examples of useful pharmaceutical adjuvants include excipients (e.g., saccharide derivatives such as lactose, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, dextrin, and carboxymethyl starch; Cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose; gum arabic; dextran; silicate derivatives such as magnesium metasilicate Aluminum; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; sulfate derivatives such as calcium sulfate, etc., binders such as gelatin, polyvinylpyrrolidone and polyethylene glycol, disintegrants For example, cellulose derivatives such as sodium carboxymethylcellulose, polyvinylpyrrolidone, lubricants (such as talc, calcium stearate, magnesium stearate, cetyl, boric acid, sodium benzoate, leucine), stabilizers ( Methylparaben, propylparaben, etc.), flavoring agents (such as commonly used sweeteners, sour agents and perfumes), diluents and solvents for injections (eg water, ethanol and glycerol, etc.) .

The dose of the compound of the present invention, a pharmaceutically acceptable salt or prodrug thereof, or a pharmaceutical composition thereof varies depending on the age, sex, race, condition, and the like of the patient.

Pharmaceutical composition and method of administration

Since the compound of the present invention has an excellent inhibitory activity against a bromodomain, the compound of the present invention and various crystal forms thereof, a pharmaceutically acceptable inorganic or organic salt, hydrate or solvate, A pharmaceutical composition containing the compound of the present invention as a main active ingredient can be used for the treatment, prevention, and alleviation of diseases associated with the activity or expression level of the Bromo domain. According to the prior art, the compounds of the present invention are useful for treating (but not limited to) various diseases such as lung cancer, bladder cancer, breast cancer, stomach cancer, liver cancer, salivary gland sarcoma, ovarian cancer, prostate cancer, cervical cancer, and epithelium. Cell carcinoma, multiple myeloma, pancreatic cancer, lymphoma, chronic myelogenous leukemia, lymphocytic leukemia, cutaneous T-cell lymphoma, etc.; T cell-mediated inflammation and autoimmune diseases such as rheumatoid arthritis, collagen II Arthritis, multiple sclerosis, systemic lupus erythematosus, psoriasis, juvenile diabetes, Sjogren's syndrome, thyroid disease, sarcoidosis, inflammatory bowel disease, celiac disease, etc. And other neurological diseases, metabolic diseases, cardiovascular diseases, viral infections, inflammation, diseases related to tissue fibrosis, and the like.

The pharmaceutical compositions of the present invention comprise a safe or effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier. By "safe and effective amount" it is meant that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. In general, the pharmaceutical compositions contain from 1 to 2000 mg of the compound of the invention per agent, more preferably from 1 to 200 mg of the compound of the invention per agent. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By "compatibility" it is meant herein that the components of the composition are capable of intermingling with the compounds of the invention and with each other without significantly reducing the efficacy of the compound. Examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid). , magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier (such as

), a wetting agent (such as sodium lauryl sulfate), a coloring agent, a flavoring agent, a stabilizer, an antioxidant, a preservative, a pyrogen-free water, and the like.

The mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration. .

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) a disintegrant such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent such as paraffin; (f) Absorbing accelerators, for example, quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, hard Calcium citrate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or a mixture thereof. In capsules, tablets and pills, the dosage form may also contain a buffer.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other materials known in the art. They may contain opacifying agents and the release of the active compound or compound in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances.

In addition to these inert diluents, the compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.

In addition to the active compound, the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.

Compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

Dosage forms for the compounds of the invention for topical administration include ointments, powders, patches, propellants and inhalants. The active ingredient is admixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or, if necessary, propellants.

The compounds of the invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When a pharmaceutical composition is used, a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight, The dose to be administered is usually from 1 to 2000 mg, preferably from 5 to 500 mg. Of course, the specific dose Factors such as the route of administration, the health of the patient, and the like should also be considered, which are within the skill of the skilled physician.

Preparation

The porphyrin derivatives according to the present invention can be prepared by various methods well known in the art of organic synthetic chemistry and those skilled in the art. The compounds of the present invention can be synthesized using the methods described hereinafter, along with synthetic methods known in the art of organic chemistry or variations as understood by those skilled in the art.

Processes for the Preparation of Compounds of Formula I of the Invention The compounds of the present invention can be prepared from readily available starting materials using the following general methods and procedures. It will be understood that when typical or preferred process operating conditions (i.e., reaction temperatures, times, moles of reactants, solvents, pressures, etc.) are given, other process operating conditions may also be used unless otherwise indicated. The optimum reaction conditions may vary depending on the particular reactants or solvents employed, but such conditions can be determined by one of ordinary skill in the art by routinely preferred procedures.

The methods of the compounds of formula I of the present invention described herein can be monitored according to any suitable method known in the art. For example, nuclear magnetic resonance, mass spectrometry, HPLC, thin layer chromatography to monitor product formation. The preparation of the compounds can involve the protection and deprotection of multiple chemical groups. The need for protection and deprotection, as well as the selection of suitable protecting groups, can be readily determined by those skilled in the art, and the chemistry of the protecting groups is in Greene and Wuts, Protective Groups in Organic Synthesis, Third Edition, Wiley & Sons, 1999.

In a preferred embodiment of the invention, the synthetic route of the compounds according to the invention is as follows:

In general, the compounds of the invention may be prepared using the reaction schemes and procedures described above, but are not limited to the reagents and solvents in the reaction conditions. R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ and R ₉ are each as defined above. Hal ₁ and Hal ₂ are halides, and the reactivity of Hal ₁ is better than that of Hal ₂ , and the reactivity of I (Hal ₁ ) is better than that of Cl (Hal ₂ ). Hal _{1 in} X can be easily converted to a phosphine-containing compound IX. For example, brominated benzene can be reacted with dimethylphosphine oxide or triethyl phosphite by Suzuki reaction to give the corresponding phosphine-containing compound. Hal _{2 in} IX can be easily converted to boric acid or borate compound VIII by the Suzuki reaction. L is a leaving functional group, such as a halogen or OH, which can be converted to a trifluoromethanesulfonic acid leaving functional group. The Suzuki reaction, such as 2,5-dibromo-3-nitropyridine and XIII, gives the pyridine compound VI. Subsequent application of a phosphine reagent, such as 1,2-bis(diphenylphosphino)ethane, to reduce the ring closure affords the phosphorus-containing porphyrin compound IV. The nitrogen in the porphyrin compound IV can be reacted with V under the reaction conditions of Mitsunobu to obtain the compound II. Then, II and intermediate compound I are subjected to a phosphine-containing porphyrin compound I under appropriate catalyst conditions.

The above-mentioned features mentioned in the present invention, or the features mentioned in the embodiments, may be arbitrarily combined. All of the features disclosed in the present specification can be used in combination with any of the compositions, and the various features disclosed in the specification can be replaced by any alternative feature that provides the same, equal or similar purpose. Therefore, unless otherwise stated, the disclosed features are only general examples of equal or similar features.

The invention is advantageous in that:

(1) The present invention provides a novel structure of a phosphine-containing porphyrin derivative;

(2) The compound provided by the present invention has a significant inhibitory effect on the Bromo region domain;

(3) The present invention provides a pharmaceutical composition for preventing or treating a disease associated with a Bromo domain domain.

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually carried out according to the conditions described in conventional conditions such as Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer. The suggested conditions. Percentages and parts are by weight unless otherwise stated.

Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those skilled in the art. In addition, any methods and materials similar or equivalent to those described may be employed in the methods of the invention. The preferred embodiments and materials described herein are for illustrative purposes only.

Preparation of Compound 1 of Example 1

Step A

Pd(OAC) 2 (7 mg, 0.03 mmol), dppf (3.3 mg, 0.06 mmol), KOAc (13 mg, 0.13 mmol) Put in a reaction bottle. After the reaction flask was closed, nitrogen was added, and THF (5 ml) was added with a syringe, followed by triethylamine (152 mg, 1.5 mmol). The mixture was stirred and heated to 68 ° C. After half a minute, a solution of triethyl phosphite (175 mg, 1.25 mmol) and compound 1a (390 mg, 1.38 mmol) in 1 ml of THF was added at 68 ° C for 4 h to dry the organic After the solvent was purified by silica gel column chromatography (EtOAc / EtOAc (EtOAc) 1.2-1.4 (m, 6H).

Step B

Add Pd(dppf) ₂ Cl _{2 to a} mixture of compound 1b (5g, 17.1mmol), boronic acid pinacol (6.5g, 25.6mmol) and KOAc (2.5g, 25.6mmol) in DMSO (50ml) (1 g), flush with nitrogen. The reaction flask was closed, and the reaction was stirred at 85 ° C overnight. After cooling to room temperature, a solution of Na ₂ CO 3 (2.5 M, 10 ml), Pd (dppf) ₂ Cl ₂ (1 g) and 2 , 5-dibromo-3-nitropyridine (7.2 g, 25.6 mmol). After 10 min of nitrogen, the reaction flask was closed and the reaction was stirred at 85 ° C overnight. The reaction solution was poured into water and extracted with ethyl acetate. the organic phase was dried with Na after mixing ₂ SO _4, drained, purified by silica gel column chromatography (PE: EA = 10: 1to 1: 1) to give a yellow solid compound 1c.

H NMR (DMSO), 8.9 (s, 1 H), 8.3 (s, 1H), 7.7-8.0 (m, 2H), 7.3-7.8 (m, 2H), 4.0-4.2 (m, 4H), 1.2-1.4 ( MS (ESI) m/z: 415 (M+H)+.

Step C

The compound 1c (5 g, 12 mmol) was stirred in EtOAc (3 mL, EtOAc (EtOAc) .HNMR (DMSO), 11.9 (s, 1H), 8.6 (s, 1H), 8.2-8.4 (m, 2H), 7.9-8.0 (m, 1H), 7.5-7.6 (m, 1H), 4.0-4.1 (m, 4H), 1.2-1.4 (m, 6H). MS (ESI) m/z: 383.0 (M+H)+.

Step D

The preparation of the intermediate a was carried out according to the method reported in the literature [Orjales, A. et al. J. Med. Chem. 2003, 46, 5512-5532 and WO2015100282]. Compound 1d (5.8 g, 10 mmol) was dissolved in DCM (100 ml). PPh3 (3.9 g, 15 mmol), Intermediate a (2.9 g, 15 mmol). The reaction mixture was stirred in ice-water bath for 1 h, then 20 ml of DIAD (3 g, 15 mmol) DCM (20 ml) was slowly added dropwise. Then, it was stirred at room temperature, and it was detected by TLC until the reaction was completed. After the solution was dried, the crude product was purified by silica gel column chromatography (DCM: MeOH=50:1 to 10:1) to give a white compound 1e.

MS (ESI) m / z: 557.0 (M+H)+.

Step E

A mixture of compound 1e (2.7 g, 5 mmol), intermediate b (2 g, 5.3 mmol), triethylamine (1.4 ml, 10 mmol), Pd (dppf) ₂ Cl ₂ (0.5 g) in DMF (30 ml) Rinse with nitrogen. After the reaction flask is closed, stir at 100-140 ° C for 4 h, pour into water. Extract with DCM. The combined organic phase is dried and evaporated to dryness. = 50: 1 to 10: 1) A yellow solid compound 1f was obtained.

H NMR (DMSO), 8.5-8.7 (m, 2H), 8.3-8.5 (m, 2H), 7.6-7.7 (m, 3H), 7.2-7.4 (m, 3H), 6.0 (d, 1H), 4.0- 4.2 (m, 7H), 3.6-4.0 (m, 2H), 3.2-3.6 (m, 2H), 2.3 (s, 3H), 1.6-1.8 (m, 2H), 1.0-1.4 (m, 9H). MS (ESI) m / z: 574.2 (M+H)+.

Step F

To a solution of the compound 1f (0.57 g, 1 mmol) in THF (10 ml), EtOAc (3 mL, EtOAc). The organic phase was combined and dried to give a crude material. EtOAcjjjjjjjjjjjjjjjjjj , 7.5-8.0 (m, 3H), 7.2-7.4 (m, 3H), 6.0 (d, 1H), 4.0 (s, 3H), 3.7-4.0 (m, 2H), 3.2-3.6 (m, 2H) , (S, 3H), 1.0-2.0 (m, 11H). MS (ESI) m/z: 514.2 (M+H)+.

Preparation of Compound 2 of Example 2

To a solution of Compound 1f (0.57 g, 1 mmol) in THF (10 mL), EtOAc (EtOAc)EtOAc. The combined organic phase was dried to give a crude material (yield: EtOAc: EtOAc:

H NMR (DMSO), 8.1-8.4 (m, 4H), 7.0-7.4 (m, 6H), 5.4 (d, 1H), 2.8-3.8 (m, 7H), 0.8-2.0 (m, 18H). ESI) m/z: 542.2 (M+H)+.

Preparation of Compound 3 of Example 3

Step A

Pd(OAC) 2 (700 mg, 3 mmol), dppf (3.3 g, 6 mmol), and KOAc (1.3 g, 13 mmol) were placed in a reaction flask. After the reaction flask was closed, nitrogen was added, and THF (100 ml) was added with a syringe, and then triethylamine (15.2 g, 150 mmol) was added. The mixture was stirred and heated to 68 ° C. After half a minute, a solution of triethyl phosphite (17.5 g, 125 mmol) and compound 3a (30 g, 138 mmol) in 5 ml of THF was added at 68 ° C for 4 h, and the organic solvent was drained. After that, silica gel column chromatography (PE / EA = 10:1 to 1:1) gave a yellow oily compound 3b.

H NMR (CDCl3), 7.5-7.7 (m, 1H), 7.2-7.3 (m, 1H), 4.2 (m, 4H), 2.3 (s, 3H), 1.3 (m, 6H). MS (ESI) m/ z: 281.7 (M+H)+

Step B

To a mixture of compound 3b (15 g, 53 mmol), boronic acid pinacol (15 g, 58.6 mmol) and KOAc (9.6 g, 98 mmol) in DMSO (200 ml) was added Pd(dppf) ₂ Cl ₂ (1.5 g) Xantphos (2g), flushed with nitrogen. The reaction flask was closed and the reaction was stirred at 110 ° C overnight. After cooling to room temperature, a solution of Na ₂ CO 3 (2.5 M, 30 mL), Pd (dppf) ₂ Cl ₂ (1 g) And 2,5-dibromo-3-nitropyridine (16.6 g, 58.6 mmol). After 10 min of nitrogen, the reaction flask was closed and the reaction was stirred at 85 ° C overnight. The reaction solution was poured into water with acetic acid The organic phase of the mixture was dried over Na ₂ SO ₄ , dried and purified by silica gel column chromatography (PE: EA = 10:1 to 1:1) to give a yellow solid compound 3c.

HNMR (CD3OD), 9.1 (s, 1H), 8.8 (s, 1H), 7.5-7.6 (m, 1H), 7.2-7.4 (m, 1H), 4.2 (m, 4H), 2.3 (s, 3H) , MS (ESI) m/z: 447.8 (M+H)+.

Step C

The compound 3c (10 g, 22.3 mmol) was stirred in EtOAc (3 mL, EtOAc (EtOAc) 3d.

HNMR (CD3OD), 8.5 (s, 1H), 8.1 (s, 1H), 8.2-8.4 (m, 2H), 7.8-8.0 (m, 1H), 4.0-4.1 (m, 4H), 2.9 (s, 3H), 1.2-1.4 (m, 6H). MS (ESI) m.

Step D

Compound 3d (5 g, 12.6 mmol) was dissolved in EtOAc (EtOAc) (EtOAcjjjjjjjjjjj 5 g, 25 mmol) DCM (20 ml) solution was added dropwise slowly, then stirred at room temperature, and then was taken up with TLC until the reaction was completed. After the solution was drained, the crude product was purified by silica gel column chromatography (DCM:MeOH = 50:1 to 10: 1) A white compound 3e was obtained.

MS (ESI) m / z: 589.2 (M+H)

Step E

A mixture of compound 3e (3g, 5.3mmol), intermediate b (4g, 10.6mmol), triethylamine (1.4ml, 10mmol), Pd(dppf) ₂ Cl ₂ (0.5g) in DMF (50ml) Rinse with nitrogen. After the reaction flask is closed, stir at 100-140 ° C for 4 h, pour into water. Extract with DCM. The combined organic phase is dried and evaporated to dryness. = 50: 1 to 10: 1) A yellow solid compound 3f was obtained. MS (ESI) m / z: 606.8 (M+H)+.

Step F

To a solution of compound 3f (1.2 g, 2 mmol) in THF (10 mL), EtOAc (EtOAc m. The mixed organic phase was dried to give a crude material. The crude product was purified by silica gel column chromatography (DCM: MeOH = 50:1 to 10:1).

HNMR (CD3OD), 8.6 (s, 1H), 8.4 (s, 1H), 8.2 (m, 1H), 7.6 (d, 2H), 7.2-7.4 (m, 3H), 5.8 (d, 1H), 4.0 (s, 3H), 4.0 (m, 1H), 3.8 (m, 1H), 3.6 (m, 1H), 3.3-3.4 (m, 2H), 3.0 (s, 3H), 2.3 (s, 3H), 1.9-2.0 (m, 7H), 1.5-1.7 (m, 1H), 1.3-1.5 (m, 1H), 1.0 (m, 1H). MS (ESI) m/z: 546.3 (M+H)+.

Preparation of Compound 4 of Example 4

Step A

Pd(OAC) 2 (700 mg, 3 mmol), dppf (3.3 g, 6 mmol), and KOAc (1.3 g, 13 mmol) were placed in a reaction flask. After the reaction flask was closed, nitrogen was added, and THF (100 ml) was added with a syringe, and then triethylamine (15.2 g, 150 mmol) was added. The mixture was stirred and heated to 68 ° C. After half a minute, a solution of triethyl phosphite (17.5 g, 125 mmol) and compound 4a (43 g, 138 mmol) in 5 ml of THF was added at 68 ° C for 4 h, and the organic solvent was drained. After that, silica gel column chromatography (PE/EA = 10:1 to 1:1) gave Compound 4b. MS (ESI) m/z: 321.7 (M+H)+

Step B

To a mixture of compound 3b (15 g, 48.8 mmol), boronic acid pinacol (15 g, 58.6 mmol) and KOAc (9.6 g, 98 mmol) in DMSO (200 ml) was added Pd(dppf) ₂ Cl ₂ (1.5 g), flush with nitrogen. The reaction flask was closed, and the reaction solution was stirred at 110 ° C overnight. After cooling to room temperature, an aqueous solution of Na ₂ CO 3 (2.5 M, 20 ml), Pd (dppf) ₂ Cl ₂ (1 g) and 2, 5-Dibromo-3-nitropyridine (16.6 g, 58.6 mmol). After 10 min of nitrogen, the reaction flask was closed and the reaction was stirred at 85 ° C overnight. The reaction solution was poured into water and extracted with ethyl acetate. after drying the organic phase with Na ₂ SO _4, drained, purified by silica gel column chromatography (PE: EA = 10: 1to 1: 1) to give a yellow solid compound 4c.

HNMR (CD3Cl), 8.9 (s, 1H), 8.5 (s, 1H), 7.8 (m, 1H), 7.0 (m, 1H), 4.0-4.3 (m, 4H), 2.6 (s, 3H), 2.0 (s, 3H), 1.2-1.4 (m, 6H). MS (ESI) m/z: 443.8 (M+H)+.

Step C

The compound 4c (10 g, 22.3 mmol) was evaporated in EtOAc (3HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 4d.

HNMR (CD3Cl), 8.9 (s, 1H), 8.6 (s, 1H), 7.8-8.0 (m, 1H), 7.6 (m, 1H), 4.1-4.2 (m, 4H), 3.0 (s, 3H) </RTI> </RTI> </RTI> <RTI ID=0.0></RTI> </RTI> <RTIgt;

Step D

Compound 4d (5 g, 12.2 mmol) was dissolved in EtOAc (EtOAc) (EtOAcjjjjjjjjjjj 5 g, 25 mmol) DCM (20 ml) solution was added dropwise slowly, then stirred at room temperature, and then was taken up with TLC until the reaction was completed. After the solution was drained, the crude product was purified by silica gel column chromatography (DCM:MeOH = 50:1 to 10: 1) A white compound 4e was obtained. MS (ESI) m / z: 585.2 (M+H)+.

Step E

A mixture of compound 4e (3 g, 5.3 mmol), intermediate b (4 g, 10.6 mmol), triethylamine (1.4 ml, 10 mmol), Pd (dppf) ₂ Cl ₂ (0.5 g) in DMF (50 ml) Rinse with nitrogen. After the reaction flask is closed, stir at 100-140 ° C for 4 h, pour into water. Extract with DCM. The combined organic phase is dried and evaporated to dryness. = 50: 1 to 10: 1) A yellow solid compound 4f was obtained. MS (ESI) m / z: 602.8 (M+H)+.

Step F

To a solution of compound 4f (1.2 g, 2 mmol) in THF (20 mL), EtOAc (EtOAc m. The mixed organic phase was dried to give a crude material. The crude material was purified by silica gel column chromatography (DCM: MeOH=50:1 to 10:1).

HNMR (CD3Cl), 8.6 (s, 1H), 7.2-7.5 (m, 7H), 6.3-6.5 (m, 1H), 4.0-4.1 (m, 1H), 3.7 (s, 3H), 3.5 (m, 1H), 2.8-3.5 (m, 6H), 2.2-2.3 (m, 6H), 2.0-2.2 (m, 7H), 0.5-1.8 (m, 4H). MS (ESI) m/z: 542.9 (M +H)+.

Preparation of Compound 5 of Example 5

Step A

Pd(OAC) 2 (700 mg, 3 mmol), dppf (3.3 g, 6 mmol), and KOAc (1.3 g, 13 mmol) were placed in a reaction flask. After the reaction flask was closed, nitrogen was added, and THF (100 ml) was added with a syringe, and then triethylamine (15.2 g, 150 mmol) was added. The mixture was stirred and heated to 68 ° C. After half a minute, a solution of triethyl phosphite (17.5 g, 125 mmol) and compound 5a (40 g, 138 mmol) in 5 ml of THF was added at 68 ° C for 4 h, and the organic solvent was drained. After that, silica gel column chromatography (PE/EA = 10:1 to 1:1) gave Compound Compound Compound Compound Compound Compound Compound Compound Compound Compound

Step B

To a mixture of compound 5b (15 g, 48.8 mmol), boronic acid pinacol (15 g, 58.6 mmol) and KOAc (9.6 g, 98 mmol) in DMSO (200 ml) was added Pd(dppf) ₂ Cl ₂ (1.5 g), flush with nitrogen. The reaction flask was closed, and the reaction solution was stirred at 110 ° C overnight. After cooling to room temperature, an aqueous solution of Na ₂ CO 3 (2.5 M, 20 ml), Pd (dppf) ₂ Cl ₂ (1 g) and 2, 5-Dibromo-3-nitropyridine (16.6 g, 58.6 mmol). After 10 min of nitrogen, the reaction flask was closed and the reaction was stirred at 85 ° C overnight. The reaction solution was poured into water and extracted with ethyl acetate. after drying the organic phase with Na ₂ SO _4, drained, purified by silica gel column chromatography (PE: EA = 10: 1to 1: 1) to give a yellow solid compound 5c.

H NMR (DMSO), 9.1 (s, 1 H), 8.9 (s, 1H), 7.5-7.8 (m, 2H), 7.4-7.5 (m, 1H), 4.0-4.2 (m, 4H), 2.2 (s, 3H), 1.1-1.3 (m, 6H). MS (ESI) m.

Step C

The compound 5c (10 g, 22.3 mmol) was refluxed in EtOAc (3 mL) (EtOAc) 5d.

HNMR (CD3Cl), 10.2 (s, 1H), 8.6 (s, 1H), 8.1 (m, 1H), 7.9-8.0 (m, 1H), 7.3 (m, 1H), 4.1-4.2 (m, 4H) , MS (ESI) m/z: 397.6 (M+H)+.

Step D

Compound 5d (5 g, 12.6 mmol) was dissolved in EtOAc (EtOAc) (EtOAcjjjjjjjjjjj 5 g, 25 mmol) DCM (20 ml) solution was added dropwise slowly, then stirred at room temperature, and then was taken up with TLC until the reaction was completed. After the solution was drained, the crude product was purified by silica gel column chromatography (DCM:MeOH = 50:1 to 10: 1) A white compound 5e was obtained. MS (ESI) m / z: 571.2 (M+H)+.

Step E

A mixture of compound 5e (3g, 5.3mmol), intermediate b (4g, 10.6mmol), triethylamine (1.4ml, 10mmol), Pd(dppf) ₂ Cl ₂ (0.5g) in DMF (50ml) Rinse with nitrogen. After the reaction flask is closed, stir at 100-140 ° C for 4 h, pour into water. Extract with DCM. The combined organic phase is dried and evaporated to dryness. = 50: 1 to 10: 1) A yellow solid compound 5f was obtained.

HNMR (CD3Cl), 8.5-8.6 (m, 1H), 8.1-8.2 (m, 1H), 7.6 (m, 1H), 7.2-7.6 (m, 6H), 5.6 (m, 1H), 4.1-4.2 ( m, 4H), 4.0-4.1 (m, 1H), 3.8-3.9 (m, 4H), 3.5-3.6 (m, 1H), 3.2-3.4 (m, 1H), 3.0-3.2 (m, 4H), 2.3 (s, 3H), 2.0 (m, 1 H), 1.0-1.8 (m, 9H). MS (ESI) m/z: 588.8 (M+H)+.

Step F

To a solution of compound 5f (1.2 g, 2 mmol) in THF (20 ml), EtOAc (EtOAc m. The mixed organic phase was dried to give a crude material (yield: EtOAc: EtOAc = 50:1 to 10:1).

HNMR (CD3Cl), 8.5-8.6 (m, 1H), 8.2-8.3 (m, 1H), 7.5-7.6 (m, 1H), 7.2-7.5 (m, 6H), 5.6-5.7 (m, 1H), 4.0-4.1 (m, 1H), 3.7-3.8 (s, 3H), 3.5-3.7 (m, 2H), 3.0-3.5 (m, 4H), 2.2-2.3 (s, 3H), 1.0-2.0 (m) , 10H). MS (ESI) m / z: 528.8 (M + H) +.

Example 6

TR-FRET BRD4 activity assay:

The BRD4 activity assay was performed according to the TR-FRET method in the Cayman kit. Dilute 3x BRD TR-FRET Assay Buffer 1 to 1x BRD TR-FRET Assay Buffer with ultrapure water. Use 1x BRD TR-FRET Assay Buffer 100 Dilute Tb-labeled donor and dye-labeled acceptor. Add 5 μl of diluted Tb-labeled donor and dye-labeled acceptor to the sample well, negative control well and positive control well. Use the configured 1x BRD TR-FRET Assay Buffer was prepared in 10% DMSO solution (the concentration of DMSO was too high to affect the reaction, the final concentration of DMSO was controlled to be 1%), and then the test compound was diluted with 10% DMSO solution. The initial screening concentration of the compound was 1 μM and 100 nM. The IC50 test starts at 10 μM or 100 μM, 3 times dilution, 8 or 10 concentration points. In addition to the control wells, add 2 μl of the diluted test compound solution to the wells used, and add 2 μl of the previously prepared 10% DMSO solution to the control wells. Dilute BET Bromodomain 40 times with 1x BRD TR-FRET Assay Buffer Ligand and Non-acetylated Ligand 1. Add 5 μl of diluted BET Bromodomain Ligand to the sample well and positive control well, and add 5 μl of diluted Non-acetylated Ligand to the negative control well 1. Use 1x BRD TR-FRET Assay Buffer BRD4 (BD1+BD2) bromodomain protein was diluted to 6 ng/μl (18 ng/well), and 3 μl of diluted BRD4 (BD1+BD2) bromodomain protein was added to all wells and mixed, and reacted at room temperature for 2 h, using ENVISION ( The fluorescence signal (320 nm stimulation, 665 nm, 615 nm emission) was detected by Perkinelmer instrument. The inhibition rate of each well was calculated from the positive control well and the negative control well. The average value of the duplicate wells was taken, and the analysis software PRISM 5.0 was used for each Compounds were tested for fit to half-inhibitory activity (IC50), Table 1.

Table 1

Compound number	FRET BRD4 IC50(nM)
1	54
2	350
3	17.3
4	8.3
5	11.2

Example 7

Cell activity assay:

The MDA-MB-231 cell line was ordered from the Shanghai Cell Resource Center of the Chinese Academy of Sciences. It was carried out according to the method in the CCK-8 kit. The logarithmic growth phase cells were collected, counted, and the cells were resuspended in complete medium to adjust the cell concentration to the appropriate level. Concentration (determined according to cell density optimization test results), inoculate 96-well plates, add 100 μl of cell suspension per well. Cells were incubated at 37 ° C, 100% relative humidity, 5% CO ₂ incubator for 24 hours. The compound was diluted to the corresponding concentration of the solution, and the cells were added at 25 μl/well. The final concentration of the compound was from 1 μM to 0 μM, and the gradient was diluted by 3 times for a total of 10 concentration points. The cells were placed at 37 ° C, 100% relative humidity, 5 Incubate for 72 hours in a %CO ₂ incubator. Add 1/10 volume of CCK-8 directly to the cell culture medium and incubate in a 37 ° C incubator for 2-4 hours. Gently shake and measure 450 nm on a SpectraMax M5 Microplate Reader. The absorbance at the wavelength was calculated by taking the absorbance at 650 nm as a reference, and the inhibition rate was calculated. The IC50 curve was fitted and the IC50 value was calculated using the software Graphpad Prism 5.

The H211 and H187 cell lines were ordered from the ATCC and assayed according to the instructions of the Alamar-Blue assay kit. The drug was dissolved in DMSO to a 50 mM stock solution and stored in a freezer at -20 °C. First, dilute the drug solution to be tested and dilute it with DMSO at a ratio of 1:3. Then dilute the DMSO-diluted drug to a final concentration of 10X in cell culture medium. The 96-well cell culture plate was taken out from the box, and 10 μl of a medium containing a series of different concentrations of the drug (10X final concentration) was added to the 96-well plate, and placed in a CO ₂ incubator for 72 hours at 37 ° C for detection, and the calculated inhibition was performed. The rate was calculated by using the GraphPad Prism 5.0 and MATILAB software using a nonlinear regression method to obtain a series of dose response curves from which the IC50 of the sample to be tested was obtained, Table 2.

Table 2

ND: No detection.

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that those skilled in the art can make various changes or modifications to the present invention after reading the above teachings of the present invention.

The scope defined by the appended claims is intended to be limited.

Claims (15)

1. A phosphine-containing porphyrin derivative and a pharmaceutically acceptable salt thereof, wherein the structure of the phosphine-containing porphyrin derivative is as shown in the following formula I:

   

   In the formula,

   R ₁ , R ₂ , R ₃ and R ₄ are each independently hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _{1 -8} alkoxy, C _1-8 alkoxycarbonyl, C _3-8 epoxyalkyl, aryl, heteroaryl, or 3- to 12-membered heterocyclic; or, adjacent R ₁ , R ₂ , R ₃ , R _{4 together} with the carbon atom to which they are bonded form a 3- to 9-membered ring;

   R _{5 is} selected from the group consisting of hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-8 alkoxy, C _1-8 An alkoxycarbonyl group, an aryl group, a heteroaryl group, or a 3- to 12-membered heterocyclic group;

   R _{6 is} selected from the group consisting of hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-8 alkoxy, C _1-8 An alkoxycarbonyl group, an aryl group, a heteroaryl group, or a 3- to 12-membered heterocyclic group;

   R _{7 is} selected from the group consisting of: hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-8 alkoxy, C _1-8 An alkoxycarbonyl group, an aryl group, a heteroaryl group, or a 3- to 12-membered heterocyclic group;

   R _{8 is} selected from the group consisting of: hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-8 alkoxy, C _1-8 Alkoxycarbonyl;

   R _{9 is} selected from the group consisting of: hydrogen, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-8 alkoxy, C _1-8 Alkoxycarbonyl;

   Wherein each of the above alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, 3- to 9-membered rings is optionally and independently substituted by one or more substituents Substituted, each of the substituents is independently hydrazine, halogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3- to 12-membered Heterocyclyl, aryl, heteroaryl, CN, NO ₂ , =O, =S.
2. The phosphine-containing porphyrin derivative according to claim 1, wherein the 3- to 9-membered ring optionally contains 1-2 additional hetero atoms selected from N, O or S.
3. The phosphine-containing porphyrin derivative according to claim 1, wherein R ₁ and R _{2 together} with the carbon atom to which they are bonded form a 3- to 9-membered ring, preferably a 5-membered ring or a 6-membered ring. .
4. The phosphine-containing porphyrin derivative according to claim 1, wherein R ₂ and R _{3 together} with the carbon atom to which they are bonded form a 3- to 9-membered ring, preferably a 5-membered ring or a 6-membered ring. .
5. The phosphine-containing porphyrin derivative according to claim 1, wherein R _{4 is} selected from the group consisting of hydrogen, halogen, C _1-3 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _1-5 alkoxy.
6. The phosphine-containing porphyrin derivative according to claim 1, wherein R _{5 is} selected from the group consisting of C _3-8 epoxyalkyl, aryl, and heteroaryl.
7. The phosphine-containing porphyrin derivative according to claim 1, wherein R _{6 is} selected from the group consisting of an aryl group, a heteroaryl group, and a 3- to 12-membered heterocyclic group.
8. The phosphine-containing porphyrin derivative according to claim 1, wherein R _{7 is} selected from the group consisting of hydrogen, halogen, C _1-3 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _1-5 straight chain and branched alkoxy group.
9. The phosphine-containing porphyrin derivative according to claim 1, wherein R _{8 is} selected from the group consisting of hydrogen, halogen, deuterated C _1-3 alkyl, C _2-4 alkenyl, C _2-4 Alkynyl; and/or

   R _{9 is} selected from the group consisting of: C _1-8 alkyl, C _1-8 haloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, C _1-5 alkoxy .
10. The phosphine-containing porphyrin derivative according to claim 1, wherein R ₈ is a deuterated methyl group.
11. The phosphine-containing porphyrin derivative according to claim 1, wherein the phosphine-containing porphyrin derivative is selected from the group consisting of:

    

    
12. A pharmaceutical composition comprising: (i) an effective amount of the compound of claim 1, or a pharmaceutically acceptable salt thereof; and (ii) pharmaceutically acceptable Carrier.
13. Use of a compound of formula I according to claim 1 or a pharmaceutically acceptable salt thereof, for use in:

    (a) preparing a Bromo domain domain inhibitor;

    (b) preparing a medicament for preventing and/or treating a disease associated with the Bromo domain; and/or

    (c) Non-therapeutic inhibition of the Bromo region domain in vitro.
14. A method of inhibiting the activity of a bromo region domain, comprising the steps of: administering an inhibitory effective amount of a compound of formula I according to claim 1 or a pharmaceutically acceptable salt thereof to a subject, or inhibiting The subject is administered an inhibitory effective amount of the pharmaceutical composition of claim 2.
15. A method of preparing a compound according to claim 1, the method comprising the steps of:

    

    (1) reacting a compound of formula II with a compound of formula III to form a compound of formula I, wherein L is a leaving group and M is a group that can be coupled to L.