Rifamycin derivatives and preparation methods and uses thereof

Abstract

The present invention relates to a rifamycin derivative represented by formula I, a method for preparing the compound and a pharmaceutical composition thereof, and the use of the compound and the pharmaceutical composition in preparing a drug for treating and / or preventing bacterial infection. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a rifamycin derivative and a preparation method and application thereof. Background Art

[0002] Rifamycins are a class of antibiotics produced by Streptococcus mediterranei. They have a broad-spectrum antibacterial effect and are very effective against Gram-positive bacteria such as Mycobacterium tuberculosis, Mycobacterium leprae, Streptococcus, Pneumococcus, and especially resistant Staphylococcus aureus. They are also effective against some Gram-negative bacteria. The rifamycin antibiotics currently used in clinical practice include rifampicin, rifapentine, and rifabutin. They are often used in combination with other anti-tuberculosis drugs to treat various types of tuberculosis and severe infections caused by resistant Staphylococcus aureus. They are also used to treat leprosy. However, due to the irrational use of antibiotics, more and more strains have developed drug resistance. The emergence of drug-resistant bacteria has increased the difficulty of curing infectious diseases and forced humans to find new ways to fight microbial infections. Summary of the invention

[0003] The present invention provides a compound represented by formula I, a pharmaceutically acceptable salt thereof, a cocrystal thereof, a tautomer thereof, a polymorph thereof, a solvate thereof, a prodrug thereof or an isotope-labeled compound thereof,

[0004]

[0005] Where: X is -N- or -CH-, R is H, C 1-6 Alkyl, amino, halogen, C 1-6 Alkylamino, di(C 1-6 Alkyl)amino or halogenated C 1-6 alkyl.

[0006] In certain embodiments, in the compound of formula I, R is H, C 1-4 Alkyl, amino, halogen, C 1-4 Alkylamino, di(C 1-4 Alkyl)amino or halogenated C 1-4 alkyl.

[0007] In certain embodiments, in the compound of formula I, R is C 1-6 Alkyl. 1-6 The alkyl group is preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl or n-hexyl.

[0008] In certain embodiments, in the compound of Formula I, R is a halogenated C 1-6 Alkyl. The halogenated C 1-6The alkyl group is preferably a halogenated methyl group, a halogenated ethyl group, a halogenated n-propyl group, a halogenated isopropyl group, a halogenated n-butyl group, a halogenated isobutyl group, a halogenated sec-butyl group, a halogenated tert-butyl group, a halogenated n-pentyl group or a halogenated n-hexyl group.

[0009] In certain embodiments, in the compound of formula I, R is halogen, preferably fluorine, chlorine, bromine or iodine.

[0010] In certain embodiments, in the compound of formula I, R is C 1-6 Alkyl, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino.

[0011] In certain embodiments, in the compound of formula I, R is C 1-4 Alkyl, amino, C 1-4 Alkylamino or di(C 1-4 Alkyl)amino.

[0012] In certain embodiments, in the compound of formula I, R is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, amino, methylamino, dimethylamino, ethylamino, diethylamino, n-propylamino or di-n-propylamino.

[0013] In certain embodiments, in the compound of formula I, R is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

[0014] In certain embodiments, in the compound of formula I, R is n-propyl, isopropyl, n-butyl or isobutyl.

[0015] In certain embodiments, in the compound of formula I, R is n-propyl or isobutyl.

[0016] In certain embodiments, in the compound of formula I, R is amino, methylamino, dimethylamino, ethylamino, diethylamino, n-propylamino or di-n-propylamino.

[0017] In certain embodiments, in the compound of formula I, R is amino, methylamino, dimethylamino, ethylamino or diethylamino.

[0018] In certain embodiments, in the compound of formula I, R is dimethylamino or diethylamino.

[0019] In certain embodiments, in the compound of Formula I, X is -N-.

[0020] In certain embodiments, in the compound of Formula I, X is -CH-.

[0021] In certain embodiments, the compound of formula I is selected from:

[0022]

[0023]

[0024] The present invention also provides a method for preparing the compound represented by formula I of the present invention, its pharmaceutically acceptable salt, its cocrystal, its tautomer, its polymorph, its solvate, its prodrug or its isotope-labeled compound, comprising:

[0025]

[0026] Compound D is reacted with a compound of formula F to generate a compound of formula I, wherein R and X are as defined in the present invention.

[0027] In certain embodiments, the method for preparing the compound of formula I of the present invention, its pharmaceutically acceptable salt, its cocrystal, its tautomer, its polymorph, its solvate, its prodrug or its isotope-labeled compound of the present invention further comprises:

[0028]

[0029] Compound C is reacted with Rifamycin S to produce Compound D.

[0030] In certain embodiments, the method for preparing the compound of formula I of the present invention, its pharmaceutically acceptable salt, its cocrystal, its tautomer, its polymorph, its solvate, its prodrug or its isotope-labeled compound of the present invention further comprises:

[0031]

[0032] Compound A is reacted with compound B to form compound C.

[0033] The present invention also provides a pharmaceutical composition, comprising a compound represented by formula I of the present invention, a pharmaceutically acceptable salt thereof, a cocrystal thereof, a tautomer thereof, a polymorph thereof, a solvate thereof, a prodrug thereof or an isotope-labeled compound thereof, and a pharmaceutically acceptable adjuvant or excipient.

[0034] The present invention also provides use of the compound represented by formula I of the present invention, its pharmaceutically acceptable salt, its cocrystal, its tautomer, its polymorph, its solvate, its prodrug or its isotope-labeled compound or the pharmaceutical composition in the preparation of a drug for preventing and / or treating bacterial infection or a disease caused by bacterial infection.

[0035] In certain embodiments, the bacterial infection is an infection caused by Gram-positive bacteria (including Mycobacterium tuberculosis, Mycobacterium leprae, Streptococcus, Pneumococcus, etc., especially drug-resistant Staphylococcus aureus) or Gram-negative bacteria (such as Pseudomonas aeruginosa).

[0036] In certain embodiments, the disease caused by the bacterial infection is tuberculosis or leprosy.

[0037] Pharmaceutical composition of the present invention can be made into various suitable dosage forms according to the route of administration. The route of administration is for example oral, parenteral, pulmonary, nasal, sublingual, tongue, buccal, rectal, percutaneous, conjunctival, topical or administered in the form of implant. The suitable dosage form includes liquid dosage form and solid dosage form, for example tablet (uncoated tablet or coated tablet, such as tablet with enteric coating or film coating), capsule, granule, oral solution, oral suspension, oral emulsion, powder, tincture, syrup, injection, suppository, ointment, cream, paste, ophthalmic preparation, pill, implant, aerosol, powder mist, spray etc. Pharmaceutical composition of the present invention or drug combination can be administered in unit dosage form. Wherein, the pharmaceutical composition, combination drug or suitable dosage form may contain 0.01 mg to 1000 mg of the compound of the present invention, its pharmaceutically acceptable salt, its cocrystal, its tautomer, its polymorph, its solvate, its prodrug or its isotope-labeled compound, preferably contains 0.1 mg to 800 mg, preferably contains 0.5-500 mg, preferably contains 0.5 to 350 mg, preferably contains 1 to 250 mg.

[0038] The compounds of the present invention as active ingredients may have systemic and / or local effects and, therefore, may be administered by a suitable route, such as oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, transdermal, conjunctival, topical or in the form of an implant.

[0039] The active ingredient can also be administered in administration forms suitable for these administration routes.

[0040] Suitable for oral administration are known administration forms which deliver the active ingredient rapidly and / or in a modified manner, such as tablets (uncoated or coated, such as tablets with enteric or mol coatings), capsules, dragees, granules, pellets, powders, emulsions, suspensions and aerosols.

[0041] With parenteral administration it is possible to avoid the absorption step (intravenous, intraarterial, intracardiac, intraspinal or intralumbar administration) or to include the absorption (intramuscular, subcutaneous, intradermal, transdermal or intraperitoneal administration). Administration forms suitable for parenteral administration. In particular, preparations in the form of solutions, suspensions, emulsions, lyophilizates and sterile powders for injection and infusion.

[0042] Suitable for other routes of administration are, for example, drugs for inhalation (especially powder inhalation, spray), nasal drops / solutions, sprays; tablets or capsules for lingual, sublingual or buccal administration, suppositories, preparations for the ears and eyes, vaginal capsules, aqueous suspensions (lotions, shaken mixtures), lipophilic suspensions, ointments, creams, emulsions, pastes, dusting powders or implants, such as stents.

[0043] The active ingredient can be converted into the administration form by methods known per se. It can be achieved with inert, non-toxic, suitable pharmaceutical excipients. It particularly includes carriers (e.g. microcrystalline cellulose), solvents (e.g. liquid polyethylene glycol), emulsifiers (e.g. sodium lauryl sulfate), dispersants (e.g. polyvinyl pyrrolidone), synthetic and natural biopolymers (e.g. proteins), stabilizers (e.g. antioxidants and ascorbic acid), colorants (e.g. inorganic pigments such as iron oxide) or flavoring agents and / or taste masking agents. In suitable cases, the active ingredient can be present in one or more of the above-mentioned carriers in the form of microencapsulation.

[0044] The pharmaceutical composition of the compound of the present invention can be administered in any of the following ways: oral, spray inhalation, rectal, nasal, vaginal, topical, parenteral, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial injection or infusion, or by means of an explanted reservoir, wherein oral, intramuscular, intraperitoneal or intravenous administration is preferred.

[0045] The compound of the present invention or the pharmaceutical composition containing the same can be administered in a unit dosage form. The dosage form can be a liquid dosage form or a solid dosage form. The liquid dosage form can be a true solution, a colloid, a microparticle dosage form, an emulsion dosage form, or a suspension dosage form. Other dosage forms include tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, freeze-dried powder injections, inclusion compounds, implants, patches, liniments, etc.

[0046] The pharmaceutical composition of the present invention may also contain commonly used pharmaceutically acceptable carriers or excipients, including but not limited to: ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycerol, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon oxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, beeswax, lanolin, etc. The content of the carrier or excipient in the pharmaceutical composition may be 1% by weight to 98% by weight, usually accounting for about 80% by weight.

[0047] Oral tablets and capsules may contain excipients such as binders, such as syrup, gum arabic, sorbitol, tragacanth, or polyvinyl pyrrolidone, fillers such as lactose, sucrose, corn starch, calcium phosphate, sorbitol, aminoacetic acid, lubricants such as magnesium stearate, talc, polyethylene glycol, silica, disintegrants such as potato starch, or acceptable wetting agents such as sodium lauryl sulfate. Tablets may be coated by methods known in pharmacy. Oral liquids may be prepared as suspensions, solutions, emulsions, syrups or elixirs of water and oil, or may be prepared as dry products, supplemented with water or other suitable media before use. Such liquid preparations may contain conventional additives, such as suspending agents, sorbitol, methyl cellulose, glucose syrup, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel, hydrogenated edible oils and fats, emulsifiers, such as lecithin, sorbitan monooleate, gum arabic; or non-aqueous carriers (which may contain edible oils), such as almond oil, fats such as glycerol, ethylene glycol, or ethanol; preservatives, such as methyl or propyl paraben, sorbic acid. Flavoring agents or coloring agents may be added as needed.

[0048] Suppositories may contain conventional suppository bases such as cocoa butter or other glycerides.

[0049] For parenteral administration, liquid dosage forms are usually made of the compound and a sterile carrier. The preferred carrier is water. Depending on the selected carrier and drug concentration, the compound can be dissolved in the carrier or made into a suspension solution. When making an injection solution, the compound is first dissolved in water, filtered and sterilized, and then filled into a sealed bottle or ampoule.

[0050] It must be recognized that the optimal dosage and interval of the compound of Formula I is determined by the properties of the compound and external conditions such as the form, route and site of administration and the specific mammal to be treated, and this optimal dosage can be determined by conventional techniques. It must also be recognized that the optimal course of treatment, i.e., the daily dosage of the compound of Formula I over a specified period of time, can be determined by methods known in the art.

[0051] The compound shown in formula I of the present invention also includes its isomers and solvates, such as hydrates, alcoholates, etc. The above-mentioned compound can also be a prodrug or can release the active ingredient in the form of metabolic changes in the body. It is a well-known technology for those skilled in the art to select and prepare appropriate prodrug derivatives. In general, for the purpose of the present invention, the solvate form with pharmaceutically acceptable solvents such as water, ethanol, etc. is comparable to the non-solvate form.

[0052] The actual dosage level of the active ingredients in the pharmaceutical composition and the combination drug of the present invention can be changed as needed to obtain the desired therapeutic effect. The dosage level must be selected according to the activity of the specific compound, the route of administration, the severity of the condition to be treated, and the condition and previous medical history of the patient to be treated. However, the conventional practice in the art is that the dosage of the compound starts from a low level and gradually increases the dosage until the desired effect is obtained. In general, the dosage of the compound of the present invention for mammals, especially humans, can be between 0.001-1000 mg / kg body weight / day, for example, between 0.01-100 mg / kg body weight / day, for example, between 0.01-20 mg / kg body weight / day.

[0053] As used herein, the term "co-crystal" refers to a complex formed by a compound of the present invention and a suitable co-crystal former through hydrogen bonding or the like. The co-crystal is preferably a physiologically acceptable co-crystal. The co-crystal former may be an organic amine, such as ethylamine, diethylamine, triethylamine, N,N′-dibenzylethylenediamine, chloroprocaine, choline, N-methylglucamine, procaine, diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, arginine, lysine or ethylenediamine. The co-crystal former may be an organic weak acid, such as fumaric acid. These co-crystals may be prepared by known co-crystal formation methods, including grinding, heating, co-sublimation, co-melting or contacting the compound of the present invention with a co-crystal former in a solution under crystallization conditions and isolating the co-crystals thus formed.

[0054] Compounds of the present invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. It is to be understood that the scope of the present invention encompasses all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0055] The present invention encompasses all possible crystalline forms or polymorphs of the compound, which may be a single polymorph or a mixture of more than one polymorph in any ratio.

[0056] The compounds of the present invention may exist in the form of solvates (e.g., hydrates), wherein the compounds of the present invention include a solvent as a structural element of the crystal lattice of the compound, such as water, methanol or ethanol. The amount of the solvent may exist in a stoichiometric or non-stoichiometric ratio.

[0057] As used in the present invention, the term "prodrug" refers to a derivative that can be hydrolyzed, oxidized or otherwise reacted under biological conditions (in vitro or in vivo) to provide the compounds of the present invention. Prodrugs become active compounds only under biological conditions through this reaction, or they have no or only low activity in their unreactive form. Prodrugs can usually be prepared using known methods, such as those described in Burger's Medicinal Chemistry and Drug Discovery (1995) 172-178, 949-982 (Manfred E. Wolff, 5th edition).

[0058] As used herein, the term "isotopically labeled compound" refers to a compound in which one or more atoms are replaced with atoms having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number predominant in nature. Examples of isotopes suitable for inclusion in the compounds of the invention include, but are not limited to, hydrogen isotopes such as 2 H, 3 H; carbon isotopes such as 11 C, 13 C and 14 C; chlorine isotopes such as 36 Cl; fluorine isotopes such as 18 F; iodine isotopes such as 123 I and 125 I; Nitrogen isotopes such as 13 N and 15 N; oxygen isotopes such as 15 Oh, 17 O and 18 O; and sulfur isotopes such as 35 S.

[0059] As used in the present invention, the term "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. Such salts include: salts formed with inorganic acids or with organic acids, such as salts formed with hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, perchloric acid, acetic acid, propionic acid, succinic acid, glycolic acid, formic acid, lactic acid, maleic acid, tartaric acid, citric acid, pamoic acid, malonic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, hydroxynaphthoic acid, hydroiodic acid, malic acid, stearic acid, tannic acid. Other acids, such as oxalic acid, although not pharmaceutically acceptable in themselves, can be used to prepare salts used as intermediates to obtain the compounds of the present invention and pharmaceutically acceptable salts thereof. Alternatively, the salt formed when the acidic proton present in the parent compound is replaced by a metal ion, such as an alkali metal ion or an alkaline earth metal ion, for example, a sodium salt, a potassium salt, a magnesium salt or a calcium salt. Alternatively, a coordination compound formed with an organic base, such as ethanolamine, diethanolamine, triethanolamine or N-methylglucamine, for example, forms an ammonium salt.

[0060] As used herein, the term "alkyl" refers to a linear or branched monovalent saturated hydrocarbon group, such as (C 1 -C 6 )alkyl refers to a group having 1 to 6 carbon atoms, such as 1, 2, 3, 4, 5 or 6 carbon atoms; (C 1 -C 4 ) Alkyl refers to a group having 1 to 4 carbon atoms, such as 1, 2, 3 or 4 carbon atoms. Non-limiting examples of alkyl include, but are not limited to, methyl, ethyl, propyl, propyl, butyl, etc.

[0061] As used in the present invention, the term "pharmaceutically acceptable carrier and / or excipient" refers to a carrier and / or excipient that is pharmacologically and / or physiologically compatible with a subject and an active ingredient, which is well known in the art (see, e.g., Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995). Pharmaceutically acceptable carriers and / or excipients include, but are not limited to, pH regulators, surfactants, ionic strength enhancers, diluents, agents that maintain osmotic pressure, agents that delay absorption, preservatives, stabilizers. For example, pH regulators include, but are not limited to, phosphate buffers. Surfactants include, but are not limited to, cationic, anionic or nonionic surfactants, such as Tween-80. Ionic strength enhancers include, but are not limited to, sodium chloride. Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, and the like. Agents that maintain osmotic pressure include, but are not limited to, sugars, NaCl, and the like. Agents that delay absorption include, but are not limited to, monostearate and gelatin. Diluents include, but are not limited to, water, aqueous buffers (such as buffered saline), alcohols and polyols (such as glycerol), etc. Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as thimerosal, 2-phenoxyethanol, parabens, chlorobutanol, phenol, sorbic acid, etc. Stabilizers have the meanings commonly understood by those skilled in the art, which can stabilize the desired activity of the active ingredient in the drug, including but not limited to sodium glutamate, gelatin, SPGA, sugars (such as sorbitol, mannitol, starch, sucrose, lactose, dextran, or glucose), amino acids (such as glutamic acid, glycine), proteins (such as dried whey, albumin or casein) or their degradation products (such as lactalbumin hydrolysate), etc.

[0062] As used herein, the term "effective amount" refers to a dose that can achieve treatment and / or prevention of the diseases or conditions described herein in a subject. The term "subject" includes humans or non-human animals. Exemplary human individuals include human individuals suffering from a disease (e.g., a disease described herein) or normal individuals. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals (e.g., birds, amphibians, reptiles), such as non-human primates, livestock and / or domesticated animals, such as dogs, monkeys, cattle, horses, etc.

[0063] As used herein, the term "treatment" is intended to alleviate, mitigate, improve or eliminate the disease state or condition being targeted. If a subject receives a therapeutic amount of the ligand-conjugated drug or its racemate, enantiomer, diastereomer, pharmaceutically acceptable salt, or a mixture of the foregoing forms according to the methods described herein, and one or more signs and symptoms of the subject show an observable and / or detectable reduction or improvement, the subject is successfully "treated". It should also be understood that the treatment of the disease state or condition includes not only complete treatment, but also failure to achieve complete treatment but achieving some biological or medically relevant results.

[0064] As used in the present invention, the term "prevention" aims to avoid, reduce, prevent or delay the occurrence of a disease or disease-related symptoms, and such disease or disease-related symptoms have not yet appeared before the administration of the relevant drug. "Prevention" does not require the complete prevention of the occurrence of a disease or disease-related symptoms. For example, after the administration of the relevant drug, the risk of a subject developing a specific disease or disease-related symptom can be reduced, or the severity of the related symptoms that appear later can be reduced, which can be considered as "preventing" the occurrence or development of the disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] Figure 1 The results show that the compounds of the examples of the present invention have an inhibitory effect on the growth of Staphylococcus aureus COL.

[0066] Figure 2 The graph shows the growth inhibition effect of the compounds of the examples of the present invention on Staphylococcus aureus USA300.

[0067] Figure 3 The results show that the compounds of the examples of the present invention have an inhibitory effect on the growth of Pseudomonas aeruginosa PAO1.

[0068] Beneficial Effects

[0069] The compound provided by the invention has an inhibitory effect on the growth of Staphylococcus aureus COL, USA300 and Pseudomonas aeruginosa PAO1. DETAILED DESCRIPTION

[0070] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. If no specific conditions are specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the raw materials, reagents or instruments used do not specify the manufacturer, they are all conventional products that can be obtained commercially.

[0071] Example 1 Preparation of Compounds

[0072] 1) Preparation of Compound C

[0073]

[0074] 40mmol of compound A was dissolved in DMF. After adding triethylamine at -15°C, 48mmol of DMF solution of compound B was added dropwise. After the addition was completed, the mixture was stirred at 25°C. The reaction was completed after 16 hours. TLC detection (PE:EA=1:2) showed that the reaction was basically complete. After the DMF was dried, an appropriate amount of water was added, and then extracted with EA. The organic phase was then washed with a saturated NaCl aqueous solution, dried, dried, and recrystallized with n-hexane. After drying, the next step of reaction was immediately carried out.

[0075] 2) Preparation of Compound D

[0076]

[0077] 11.7 mmol of compound C was dissolved in toluene, and 11.7 mmol of Rifamycin S was added under stirring at room temperature. The reaction solution was black, and the reaction was basically completed after 16 hours. The toluene was dried by spin drying, and then 95% ethanol was added until it was completely dissolved. MnO 2 After stirring at room temperature for 0.5 hours, the plate was spotted (PE:EA=1:1), indicating that the reaction was basically completed. After being spin-dried, it was purified by chromatographic column and then immediately proceeded to the next step of reaction.

[0078] 3) Preparation of compound DYC-2022-02

[0079]

[0080] DMSO was added to a single-necked bottle containing 4.6 mmol of compound D, and 46 mmol of MnO was added under stirring. 2 , 4.6mmol of compound E was drawn into the system by syringe, and the resulting system was black, and then turned dark green. The reaction was carried out at 40℃ for 14 hours, and TLC monitoring (PE:EA=1:1) showed that product spots were generated, and the raw materials were basically reacted; EA was added to the reaction system for dilution, filtered, and the organic phase was washed with water, and the aqueous phase was back-extracted with EA, the organic phases were combined, dried and spin-dried to obtain a blue solid, which was purified by chromatographic column to obtain a relatively pure compound DYC-2022-02, totaling 2.5g.

[0081] 1H NMR(500MHz,Chloroform-d)δ9.47(s,1H),8.26(s,1H),7.28-7.18(m,1H),7.08(ddq,1H),6.44(d,1H),6.38(dd,1 H),6.32(d,1H),6.09-5.99(m,1H),5.14(ddd,1H),5.02-4.94(m,1H),4.41(d,1H),3.83(d,1H),3.77-3.61(m,2H) ,3.46(ddd,2H),3.34(ddddd,1H),3.30-3.23(m,5H),3.04(ddtq,1H),2.45-2.34(m,1H),2.24(d,6H),2.13-2.03( m,8H),1.87-1.60(m,7H),1.23(tqd,1H),1.04(dt,3H),0.94-0.86(m,6H),0.81(dt,3H).MS(ESI)m / z:927.43[M+H] + ; 949.45[M+Na] + .

[0082] Use Substituting compound E, refer to step 3) to prepare compounds DYC-2022-02 and DYC-2022-03.

[0083]

[0084] 1 H NMR(500MHz,Chloroform-d)δ9.47(s,1H),8.26(s,1H),7.28-7.18(m,1H),7.08(ddq,1H),6.44(d,1H),6.38(dd,1H), 6.32(d,1H),6.09-5.99(m,1H),5.14(ddd,1H),4.98(dddd,1H),4.41(d,1H),3.83(d,1H),3.77-3.61(m,2H),3.34(td p,1H),3.29-3.23(m,7H),2.68-2.62(m,4H),2.40(dddt,1H),2.33(d,2H),2.16(s,3H),2.08-2.03(m,6H),1.79-1.68 (m,5H),1.68-1.60(m,1H),1.23(tqd,1H),1.04(dt,3H),0.94-0.84(m,12H),0.81(dt,3H).MS(ESI)m / z:941.44[M+H] +; 963.45[M+Na] + .

[0085]

[0086] 1 H NMR(500MHz,Chloroform-d)δ9.47(s,1H),8.26(s,1H),7.28-7.18(m,1H),7.08(ddq,1H),6.44(d,1H),6.38(dd ,1H),6.32(d,1H),6.09-5.99(m,1H),5.14(ddd,1H),4.98(dddd,1H),4.41(d,1H),3.83(d,1H),3.77-3.61(m,2H ),3.34(tdp,1H),3.30-3.25(m,7H),2.78-2.73(m,4H),2.44(t,2H),2.39(dddt,1H),2.16(s,3H),2.08-2.03(m, 6H),1.79-1.51(m,7H),1.23(tqd,1H),1.04(dt,3H),0.94-0.85(m,9H),0.81(dt,3H).MS(ESI)m / z:927.43[M+H] + ; 949.45[M+Na] + .

[0087] Example 2 Evaluation of in vitro killing effect on Staphylococcus aureus

[0088] CCK8 to measure cell viability and make IC 50 curve.

[0089] Pick a single clone of Staphylococcus aureus USA300 or COL bacteria (purchased from ATCC) grown on BHI (BD) plates and introduce it into BHI (BD) medium, culture it at 37°C, 220rpm, and culture it overnight; centrifuge it at 4000rpm in a normal temperature centrifuge for 5 minutes, collect the bacteria, wash them 3 times with sterile PBS solution, measure OD600, and use culture medium to adjust the initial bacterial density to OD600 of 0.1; dilute the antibiotics by 2 times from the highest 5μg / mL, add 100μL of the diluted antibiotics and an equal volume of bacteria with OD600 of 0.1 to a 96-well plate, mix them thoroughly, and culture them in a constant temperature incubator at 37°C for 24 hours, measure OD600 and calculate the viability. The results are shown in Tables 1 and 2, and Figure 1 and Figure 2 .

[0090] Table 1 IC values ​​of the compounds of the present invention against Staphylococcus aureus COL 50 concentration

[0091]

[0092] Table 2 IC values ​​of the compounds of the present invention against Staphylococcus aureus USA300 50 concentration

[0093]

[0094] Example 3 Evaluation of in vitro killing effect on Pseudomonas aeruginosa

[0095] Pseudomonas aeruginosa PAO1 (purchased from ATCC) grown on LB plates was picked and introduced into LB medium, cultured overnight at 37°C, 220rpm; centrifuged at 4000rpm for 5 minutes in a normal temperature centrifuge, the bacteria were collected, washed 3 times with sterile PBS solution, OD600 was measured, and the initial bacterial density was adjusted to OD600 of 0.1 with culture medium; the antibiotics were diluted 2 times from the highest 100μg / mL, and 100μL of the diluted antibiotics and an equal volume of bacteria with OD600 of 0.1 were added to a 96-well plate, mixed thoroughly, and cultured in a 37°C constant temperature incubator for 24 hours, and OD600 was measured and the viability was calculated. The results are shown in Tables 3 and Figure 3 .

[0096] Table 3 IC values ​​of the compounds of the present invention against Pseudomonas aeruginosa PAO1 50 concentration

[0097]

[0098] The above experimental results show that the compounds provided by the present invention have an inhibitory effect on the growth of Staphylococcus aureus COL, USA300 and Pseudomonas aeruginosa PAO1.

[0099] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, ordinary technicians in the field should understand that the specific implementation methods of the present invention can still be modified or some technical features can be replaced by equivalents without departing from the spirit of the technical solution of the present invention, which should be included in the scope of the technical solution for protection of the present invention.

Claims

1. Use of compound DYC-2022-03 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing compound DYC-2022-03 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating bacterial infection or a disease caused by bacterial infection, The bacterial infection is an infection caused by Pseudomonas aeruginosa, and the disease caused by the bacterial infection is a disease caused by Pseudomonas aeruginosa infection.

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