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WO2016095830A1 - Conjugué peptide-triterpène pentacyclique, et procédé de préparation, composition pharmaceutique et utilisation de celui-ci - Google Patents

Conjugué peptide-triterpène pentacyclique, et procédé de préparation, composition pharmaceutique et utilisation de celui-ci Download PDF

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Publication number
WO2016095830A1
WO2016095830A1 PCT/CN2015/097685 CN2015097685W WO2016095830A1 WO 2016095830 A1 WO2016095830 A1 WO 2016095830A1 CN 2015097685 W CN2015097685 W CN 2015097685W WO 2016095830 A1 WO2016095830 A1 WO 2016095830A1
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pharmaceutically acceptable
acid
acceptable salt
conjugate
group
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Chinese (zh)
Inventor
刘克良
娜荷芽
李相鹏
王晨宏
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Institute of Pharmacology and Toxicology of AMMS
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Institute of Pharmacology and Toxicology of AMMS
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
    • C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents

Definitions

  • the invention belongs to the field of medicine and chemical industry, and relates to a pentacyclic triterpene-peptide conjugate, a preparation method thereof, a pharmaceutical composition and use thereof.
  • Antimicrobial peptides have broad-spectrum bactericidal properties and are resistant to microorganisms such as bacteria, fungi, protozoa and viruses.
  • the antibacterial peptide first discovered by Swedish scientists in the silkworm pupa in 1980. At present, scientists from various countries have discovered more than 2,000 different kinds of antimicrobial peptides. Although the amino acid sequences, peptide chain lengths and structures of these antimicrobial peptides are different, different kinds of antimicrobial peptides have the following common characteristics:
  • the antimicrobial peptide is formed by condensation of 10-50 amino acids
  • the antimicrobial peptide is generally a cationic peptide with 2-9 positive charges
  • the antimicrobial peptide contains about 50% of non-polar amino acid residues, and is both hydrophilic and hydrophobic.
  • the cationic character of the antimicrobial peptide and the hydrophilic and hydrophobic amphiphilic properties allow the antimicrobial peptide to adhere to the anionic component on the surface of the bacterial cell membrane by electrostatic interaction, thereby promoting the hydrophobic group to be sterilized by hydrophobic interaction or van der Waals force embedded in the bacterial cell membrane. Effectiveness. Lysine and arginine are basic amino acids that are used frequently in the composition of antimicrobial peptides.
  • Pentacyclic triterpenoids which have a wide range of pharmacological effects and important biological activities, especially showing pharmacological properties of interest in anti-inflammatory and antibacterial, skin care, anti-tumor and immune regulation.
  • Rosenei L. Brum and other five-ring triterpenoids Betulinic acid, isolated from the skin of the candle tree (Vochysia divergens) inhibited Staphylococcus aureus; G.P.P.Kamatou et al., which were isolated from 16 species of South African sage, Oleanolic acid and Ursolic acid, showed activity by antibacterial activity.
  • the inventors of the present invention creatively combine antibacterial peptides with pentacyclic triterpenoids to design a novel structure of the conjugate.
  • the active compounds show that the new compounds have excellent antibacterial activity, and the minimum inhibitory concentration can even reach Several ⁇ M levels.
  • One aspect of the present invention provides a pentacyclic triterpene-peptide conjugate represented by the formula (I) or a pharmaceutically acceptable salt thereof,
  • XA is a pentacyclic triterpenoid or an esterified derivative thereof;
  • P is a polypeptide having a length of 2 to 50 amino acids and containing one or more arginine and/or lysine; The C terminus is a free carboxyl group or amidation;
  • L is a tether.
  • the pentacyclic triterpene-peptide conjugate or a pharmaceutically acceptable salt thereof wherein the content of arginine and/or lysine in the polypeptide is ⁇ 50 %, ⁇ 60%, ⁇ 70%, ⁇ 80%, ⁇ 90% or 100%; preferably, the arginine is L-arginine; preferably, the lysine is L-lysine acid.
  • the pentacyclic triterpene-peptide conjugate or a pharmaceutically acceptable salt thereof wherein the polypeptide is 2-40 amino acids in length, 2-30 amino acids in length, 2 -20 amino acids, 2-15 amino acids, 2-12 amino groups Acid, 2-10 amino acids, 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids, 8 amino acids, 9 amino acids or 10 amino acids; preferably, said The amino acid is L-arginine or L-lysine.
  • the polypeptide is an antimicrobial peptide.
  • the polypeptide is a non-antibacterial peptide.
  • the pentacyclic triterpene-peptide conjugate or a pharmaceutically acceptable salt thereof wherein the linker L is azidoacetic acid; preferably, the azide acetate passes through a carboxyl group Attached to the ⁇ -amino group at the N-terminus of the polypeptide, the azide group of azide acetate is linked to the alkynyl group of XA via a Husigen cycloaddition reaction; preferably, the Husigen cycloaddition reaction is carried out under cuprous ion catalysis.
  • the pentacyclic triterpene-peptide conjugate or a pharmaceutically acceptable salt thereof wherein the esterified derivative is an esterified derivative of a pentacyclic triterpenoid
  • the esterification site is a 3-hydroxyl group and/or a 28-position carboxyl group of a pentacyclic triterpenoid; more preferably, the esterified derivative contains an alkynyl group; further preferably, betulinic acid, olean
  • the 28-position carboxyl group of acid and ursolic acid is esterified to obtain alkyn-containing BAo, OAo and UAo (see below for structural formula), and the esterification reaction of betulinic acid, oleanolic acid and ursolic acid at the 3 position hydroxyl group respectively
  • the alkynyl-containing BAc, OAc and UAc are obtained (see the structural formula below).
  • the pentacyclic triterpene-peptide conjugate or a pharmaceutically acceptable salt thereof is selected from the group consisting of conjugates or pharmaceutically acceptable salts thereof:
  • L is azidoacetic acid and R is L-arginine.
  • RRRR is represented by SEQ ID NO: 1
  • RRRRRR is represented by SEQ ID NO: 2
  • RRRRRR is represented by SEQ ID NO: 3
  • RRRRRRR is represented by SEQ ID NO: 4
  • RRRRRRRR is represented by SEQ ID NO: 5.
  • the bacterium is a Gram-positive bacterium;
  • the Gram-positive bacteria are at least one selected from the group consisting of Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus cereus.
  • the conjugate mentioned hereinafter refers to the pentacyclic triterpene-peptide conjugate of the present invention, unless otherwise specified.
  • Another aspect of the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising the pentacyclic triterpene-peptide conjugate of any of the above and/or a pharmaceutically acceptable salt thereof; optionally, further comprising a pharmaceutically acceptable Acceptable excipients (eg, carriers or excipients, etc.).
  • the pharmaceutical composition consists of a pentacyclic triterpene-peptide conjugate of the invention and/or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable adjuvant.
  • the pentacyclic triterpene-peptide conjugate of the present invention and/or a pharmaceutically acceptable salt thereof is used as an active ingredient.
  • the pharmaceutical compositions of the invention contain from 0.1 to 90% by weight of the conjugate and/or a pharmaceutically acceptable salt thereof.
  • Pharmaceutical compositions can be prepared according to methods known in the art.
  • the conjugate or a pharmaceutically acceptable salt thereof can be combined with one or more solid or liquid pharmaceutical excipients to provide a suitable administration form or dosage form for human use.
  • the conjugate of the present invention or the pharmaceutical composition containing the same may be administered in a unit dosage form, which may be enterally or parenterally, such as oral, muscle, subcutaneous, nasal, oral mucosa, skin, peritoneum or rectum.
  • a unit dosage form which may be enterally or parenterally, such as oral, muscle, subcutaneous, nasal, oral mucosa, skin, peritoneum or rectum.
  • Formulations such as tablets, capsules, pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, buccal tablets, suppositories, lyophilized powders Wait. It may be a general preparation, a sustained release preparation, a controlled release preparation, and various microparticle delivery systems.
  • various excipients known in the art can be widely used.
  • carriers are, for example, diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline fibers.
  • diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline fibers.
  • wetting agent, aluminum silicate, etc. wetting agent and binder, such as water, glycerin, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, gum arabic, gelatin pulp, carboxy Methylcellulose sodium, shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.; disintegrating agents, such as dried starch, alginate, agar powder, brown algae starch, sodium hydrogencarbonate and tannic acid, calcium carbonate , polyoxyethylene, sorbitan fatty acid ester, sodium dodecyl sulfate, methyl cellulose, ethyl cellulose, etc.; disintegration inhibitors, such as sucrose, glyceryl tristearate, cocoa butter, hydrogenation An oil or the like; an absorption enhancer such as a quaternary ammonium salt, sodium lauryl sulfate or the like; a lubricant such as
  • Tablets may also be further formed into coated tablets, such as sugar coated tablets, film coated tablets, enteric coated tablets, or bilayer tablets and multilayer tablets.
  • various carriers known in the art can be widely used.
  • the carrier are, for example, diluents and absorbents such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, kaolin, talc, etc.; binders such as acacia, tragacanth, gelatin , ethanol, honey, liquid sugar, rice paste or batter; etc.; disintegrating agents, such as agar powder, dried starch, alginate, sodium dodecyl sulfate, methyl cellulose, ethyl cellulose, and the like.
  • the active ingredient conjugate or a pharmaceutically acceptable salt thereof is mixed with the various carriers described above, and the resulting mixture is placed in a hard gelatin capsule or soft capsule.
  • the active ingredient conjugate or a pharmaceutically acceptable salt thereof may also be formulated as a microcapsule, suspended in an aqueous medium to form a suspension, or may be enclosed in a hard capsule or used as an injection.
  • an injection preparation such as a solution, an emulsion, a lyophilized powder injection and a suspension
  • all diluents conventionally used in the art for example, water, ethanol, polyethylene glycol, 1, 3 may be used.
  • an appropriate amount of sodium chloride, glucose or glycerin may be added to the preparation for injection, and a conventional solubilizer, a buffer, a pH adjuster or the like may be added.
  • coloring agents may also be added to the pharmaceutical preparations as needed.
  • the dosage of the conjugate of the present invention or a pharmaceutically acceptable salt thereof depends on a number of factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, weight and individual response of the patient or animal, and the specific conjugation used.
  • the above dosages may be administered in a single dosage form or divided into several, for example two, three or four dosage forms.
  • composition as used herein is intended to include a product comprising specified amounts of each of the specified ingredients, as well as any product that results, directly or indirectly, from the specified combination of the specified ingredients.
  • each active ingredient in the pharmaceutical compositions of the present invention can be varied so that the amount of active ingredient is effective to provide the desired therapeutic response to the particular patient, composition, and mode of administration.
  • the dosage level will be selected based on the activity of the particular conjugate, the route of administration, the severity of the condition being treated, and the condition and past medical history of the patient to be treated. However, it is the practice in the art that the dosage of the conjugate begins at a level below that required to achieve the desired therapeutic effect, gradually increasing the dosage until the desired effect is achieved.
  • a further aspect of the present invention provides a process for the preparation of the pentacyclic triterpene-peptide conjugate, comprising the steps of: esterifying a derivative of a pentacyclic triterpenoid with a polypeptide-azidoacetate condensate in a cuprous The Hsigen cycloaddition reaction is carried out under ion catalysis to obtain a final product; wherein the esterified derivative of the pentacyclic triterpenoid is obtained by esterification of a pentacyclic triterpenoid; the polypeptide-azidoacetic acid
  • the condensate is prepared by azidoacetic acid condensed by a carboxyl group and an ⁇ -amino group at the N-terminus of the polypeptide.
  • the pentacyclic triterpenoid has a double bond inside/outside the ring.
  • a pentacyclic triterpenoid or a pentacyclic triterpenoid compound using an aminoalkanoic acid as a tether is directly reacted with a peptide resin by a solid phase synthesis method in a general sense, a five-ring triterpene under acid cracking (TFA) conditions
  • TFA acid cracking
  • the double bond will shift or loop and will not form a pentacyclic triterpene-peptide conjugate.
  • pentacyclic triterpenoids cannot be directly linked to the cleaved peptide resin.
  • the polypeptide-azidoacetic acid modification is prepared by the following steps: Fmoc solid phase synthesis to obtain an amino terminal exposed peptide resin, and then blocking the exposed amino end with azide acetic acid The blocked peptide resin is cleaved and purified to obtain a polypeptide-azidoacetate modification.
  • azide acetic acid, HBTU, HOBT and DIEA are added to the peptide resin exposed at the amino terminal end during blocking, and the reaction is carried out at room temperature, preferably for 1 hour.
  • the lysate is added to the blocked peptide resin during the cleavage, and the reaction is first carried out in an ice bath and then at room temperature; preferably, the reaction is carried out for 30 minutes in an ice bath and then at room temperature for 120 minutes.
  • the lysate used for the cleavage consists of TFA, ethanedithiol, anisole and water; preferably, from 90% v/v TFA, 5% v/v ethane disulfide Alcohol, 2.5% v/v anisole and 2.5% v/v water; more preferably, TFA is pre-ice bathed for 30 min or pre-stored in the refrigerator for later use.
  • the purification is carried out by medium pressure liquid chromatography or high pressure liquid chromatography; preferably, the eluent used is acetonitrile, water and a small amount (0.1% v/v) of trifluoroacetic acid. .
  • a further aspect of the invention provides the pentacyclic triterpene-peptide conjugate or a pharmaceutically acceptable salt thereof or the pharmaceutical composition for preparing an antibacterial or therapeutic And/or use in a medicament for preventing and/or adjuvant treatment of a disease caused by a bacterial infection or a bacterial infection; preferably, the bacteria or bacteria are Gram-positive bacteria; preferably, the Gram-positive bacteria are selected from At least one of Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus cereus.
  • the present invention also provides a method of antibacterial in vivo or in vitro comprising the step of using an effective amount of a pentacyclic triterpene-peptide conjugate of the present invention or a pharmaceutically acceptable salt thereof; preferably, the bacterium is Gram-positive bacteria; more preferably, the Gram-positive bacteria are at least one selected from the group consisting of Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus cereus.
  • a further aspect of the invention relates to a method of treating and/or preventing and/or adjunctively treating a disease caused by a bacterial infection or a bacterial infection comprising the use of an effective amount of a pentacyclic triterpene-peptide conjugate of the invention or a pharmaceutical thereof a step of accepting a salt;
  • the bacterium is a Gram-positive bacterium; more preferably, the gram-positive bacterium is selected from the group consisting of Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, and waxy At least one of Bacillus.
  • a therapeutically and/or prophylactically effective amount of a conjugate of the invention may be applied in pure form or in the form of a pharmaceutically acceptable salt.
  • the conjugate can be administered as a pharmaceutical composition comprising the conjugate of interest and one or more pharmaceutically acceptable excipients. It will be appreciated, however, that the total daily usage of the conjugates and compositions of the invention will be determined by the attending physician within the scope of sound medical judgment.
  • the specific therapeutically effective dosage level for any particular patient will depend on a number of factors, including the disorder being treated and the severity of the disorder; the activity of the particular conjugate employed; the particular combination employed.
  • the dosage of the conjugate begins at a level below that required to achieve the desired therapeutic effect, gradually increasing the dosage until the desired effect is achieved.
  • the dosage of the conjugates of the invention for use in mammals, particularly humans may range from 0.001 to 1000 mg/kg body weight per day, for example between 0.01 and 100 mg/kg body weight per day, for example between 0.01 and 10 mg/kg. Weight / day.
  • the pentacyclic triterpene-peptide conjugate according to the present invention or a pharmaceutically acceptable salt thereof can effectively prevent and/or treat various diseases or conditions described in the present invention.
  • the term "effective amount” refers to a dose that can achieve a treatment, prevention, alleviation, and/or alleviation of a disease or condition described herein in a subject.
  • disease and/or condition refers to a physical state of the subject that is associated with the disease and/or condition described herein.
  • subject can refer to a patient or other animal that receives the pharmaceutical composition of the invention to treat, prevent, ameliorate and/or alleviate the disease or condition of the invention, particularly a mammal, such as a human, a dog, a monkey, or a cow. , horses, etc.
  • Fmoc solid phase synthesis means, unless otherwise specified, a conventional 9-fluorenylmethoxycarbonyl (Fmoc) solid phase polypeptide synthesis method.
  • Humansigen cycloaddition reaction means: copper-catalyzed azide-alkyne Husigen cycloaddition reaction (Copper-Catalyzed Azide-Alkyne Cycloaddition).
  • the pentacyclic triterpene-peptide conjugate of the present invention has antibacterial and antibacterial activity, and has excellent antibacterial activity especially for Gram-positive bacteria, and can inhibit infection caused by Gram-positive bacteria.
  • TFA Trifluoroacetic acid-trifluoroacetic acid
  • MIC Minimum Inhibitory Concentration
  • MALDI-TOF-MS-matrix assisted laser desorption-tandem time-of-flight mass spectrometry
  • the solid phase synthesis carrier Rink amide resin used in the following preparation examples/examples was provided by Tianjin Nankai Synthetic Co., Ltd.; HBTU, HOBT, DIEA and Fmoc protected natural amino acids were provided by Shanghai Jill Biochemical Co., Ltd.; Trifluoroacetic acid (TFA) was Beijing Products of Bomaijie Technology Co., Ltd.; DMF and DCM are products of Sinopharm Chemical Reagent Co., Ltd.; chromatographic pure acetonitrile is Fisher's product. Other reagents are domestically produced pure products if they are not described.
  • R represents L-arginine
  • the synthetic route is as follows:
  • the solid phase synthesis reactor was bubbled, washed, thoroughly dried, and then soaked overnight by adding a silylating agent (an anhydrous toluene solution containing 10% v/v trimethylchlorosilane).
  • a silylating agent an anhydrous toluene solution containing 10% v/v trimethylchlorosilane.
  • the silylation reagent is recovered, and the reactor is washed with an anhydrous organic solvent and dried for use.
  • step 2 a peptide resin of a certain length is obtained, and then the azide acetic acid is condensed at the amino terminus, and the specific steps are as follows:
  • the peptide resin was impregnated with 25% v/v piperidine in DMF solution to remove the Fmoc protecting group ⁇ the resin was washed with DMF, MeOH and DCM in turn ⁇ the ninhydrin reagent was detected positive ⁇ the azide acetic acid, HBTU, HOBT were put into the peptide resin. The reaction was stirred for 1 hour at room temperature with DIEA.
  • the peptide resin was washed three times with DMF, shrunk with anhydrous methanol, and dried under vacuum at room temperature.
  • the peptide resin was placed in a 250 mL eggplant-shaped flask, and the lysate was added in an ice bath under electromagnetic stirring at a ratio of 10 mL/g of peptide resin.
  • the lysate consists of 90% v/v TFA, 5% v/v ethanedithiol, 2.5% v/v anisole and 2.5% v/v water.
  • TFA needs to be cooled in advance by ice bath for 30 min or in advance. Store in the refrigerator for later use. After adding the lysate to the ice bath, the resin turned orange-red. After reacting for 30 minutes, the ice bath was removed, and the reaction was continued at room temperature for 120 minutes and then ended.
  • the crude product obtained was purified by medium pressure liquid chromatography or high pressure liquid chromatography, and the column was a C8 column.
  • the eluent was acetonitrile, water and a small amount (0.1% v/v) trifluoroacetic acid. Specific procedure: Weigh 0.5g of crude product, add 20mL of water to dissolve the solid, centrifuge for 10min (3000r/min) and take the supernatant for loading.
  • the column is pre-treated with 5% acetonitrile/water/0.1% trifluoroacetic acid solution (volume percentage) 160mL balance, after sample loading, continue to rinse with 5% acetonitrile / water / 0.1% trifluoroacetic acid solution (volume percent) 200mL, high performance liquid chromatography to detect eluent components, if no peptide-azidoacetate modification is detected
  • the main peak needs to gradually increase the acetonitrile content until the main peak is eluted.
  • the eluates were combined, and most of the solvent was removed by rotary evaporation, and the pure peptide-azidoacetic acid modified product was freeze-dried, and the purity was more than 95% by HPLC.
  • Substituting compound 1 betulinic acid for oleanolic acid or ursolic acid (same reaction route, correspondingly preparing OAo or UAo), and/or extending the peptide sequence to RR, RRR, RRRRRR or RRRRRRRR (according to the procedure of Preparation Example 1) 2)
  • the solid phase synthesis method of a) achieves the extension of the peptide sequence) to obtain pentacyclic triterpene-peptide conjugates (2)-(3), (7)-(9), (13)-(15), (19)-(21), (25)-(27).
  • the synthetic route is as follows:
  • the synthesis process is the same as that of the pentacyclic triterpene-peptide conjugate (1) except that BAc is replaced with BAo.
  • the preparation method is referred to Preparation Example 16.
  • the preparation of the polypeptide is 4 arginine, 5 arginine or 7 arginine. Conjugate.
  • Gram-positive bacteria Bacillus subtilis (B. subtilis), Staphyloccus aureus (S. aureus), Staphyloccus epidermids (S. epidermids), Bacillus cereus , B.cereus).
  • Gram-negative bacteria Escherichia coli (E. coli), Pseudpmonas aeruginosa (P. aeruginosa), Serratia marcescens (S. marcescens).
  • Bacterial solution E. coli glycerol suspension, Pseudomonas aeruginosa glycerol suspension, Serratia marcescens glycerol suspension, Bacillus subtilis glycerol suspension, Staphylococcus aureus glycerin suspension, Staphylococcus epidermidis glycerol suspension and waxy sample Bacillus glycerol suspension Inoculum, 80% (v/v) glycerol suspension, stored at -80 °C.
  • Nutritional broth 3g beef extract, 10g peptone and 5g NaCl were dissolved in 800mL of water, adjusted to pH 7.2-7.4 with KOH solution, made up to 1000mL with water, autoclaved at 121 °C for 20-30min.
  • Negative control nutrient broth without added drugs.
  • Sample Pelican, betulinic acid, oleanolic acid, ursolic acid, peptides with N-terminal azide acetate (N 3 -RRR and N 3 -RRRRRRRR), and pentacyclic triterpenes in the above preparation examples - Peptide conjugates (13)-(15), (16)-(18), (28)-(30) were used as samples, and about 1 mg of the sample was weighed, and a 1 mg/mL solution was prepared by using a sterile injection solution. As a sample solution (adjust the concentration according to the activity).
  • the number of samples take the appropriate number of 96-well plates (12 holes per row, 8 lines in total; Costar 3799, Corning Incorporation, USA), first add 100 ⁇ L of nutrient broth to each well, then add in the first row of wells. The sample solution was added in an amount of 100 ⁇ L/well, and each sample was repeated in three rows along the same row, so that a total of 4 samples were arranged in the first row of wells (a positive control was included in the sample).
  • step-by-step dilution method use the multiple dilution method (step-by-step dilution method) to operate, that is, first fully mix the solution in the first row of holes, then use the lance to absorb 100 ⁇ L and add to the second row of holes to mix, then take the second row of holes 100 ⁇ L of solution inside Go to the third row of holes, and so on to the last row, dilute to the last row and then take 100 ⁇ L and discard.
  • three wells in the lower right corner of the 96-well plate contained only nutrient broth as a negative control.
  • the clarification of the wells in the 96-well plate was taken out, and the OD 600 value of each well was measured by a microplate reader, and the bacterial growth rate of each well was calculated according to the following formula.
  • the clarification in the well was observed, no significant turbidity, and the lowest concentration of bacterial growth rate ⁇ 50% in the calculated pores was the minimum inhibitory concentration (MIC) of the sample.
  • Bacterial growth rate (OD 600 sample / OD 600 negative control ) ⁇ 100%
  • the pentacyclic triterpene-peptide conjugate of the present invention exhibits good antibacterial activity against bacteria, particularly Gram-positive bacteria, and the minimum inhibitory concentration is at several micromolar levels,
  • the positive control, phlorizan is comparable and far superior to the corresponding pentacyclic triterpenoids or short peptides alone.
  • Negative control nutrient broth without added drugs.
  • the conjugate of the present invention has good inhibitory activity against S. aureus, B. subtilis and E. coli, especially for Staphylococcus aureus. And B. subtilis, the minimum inhibitory concentration is at several micromolar levels, comparable to the positive control Pelican.
  • the results also showed that the number of arginine is equal to 3, and its modification site is at 28-COOH. (ie, compound BAo-RRR, OAo-RRR, UAo-RRR) is the best against E. coli growth activity.

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Abstract

La présente invention concerne un conjugué peptide-triterpène pentacyclique, et un procédé de préparation, une composition pharmaceutique et une utilisation de celui-ci. Ledit conjugué peptide-triterpène pentacyclique présente un effet antibactérien et, en particulier, un importante effet antibactérien sur les bactéries à gram-positif.
PCT/CN2015/097685 2014-12-18 2015-12-17 Conjugué peptide-triterpène pentacyclique, et procédé de préparation, composition pharmaceutique et utilisation de celui-ci Ceased WO2016095830A1 (fr)

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CN201410789419.XA CN105777864B (zh) 2014-12-18 2014-12-18 五环三萜-肽缀合物、其制备方法、药物组合物及用途

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11266632B1 (en) * 2021-07-20 2022-03-08 King Abdulaziz University Maslinic and oleanolic acids derivatives for treating SARS-CoV-2 infection
CN119431488A (zh) * 2024-11-05 2025-02-14 西北大学 齐墩果烷型三氮唑糖苷化合物及其在制备ptp1b抑制剂和抗糖尿病药物中的应用

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* Cited by examiner, † Cited by third party
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CN113975404B (zh) * 2021-09-22 2023-06-20 中国农业科学院兰州畜牧与兽药研究所 一种氟苯尼考多肽衍生物及其应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060166867A1 (en) * 2003-04-28 2006-07-27 Yeda Research And Development Co. Ltd. Novel conjugates of polysaccharides and uses thereof

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1682740A (zh) * 2005-03-11 2005-10-19 中国药科大学 五环三萜类化合物在制备糖原磷酸化酶抑制剂中的应用
CN101580530A (zh) * 2008-05-14 2009-11-18 北京美倍他药物研究有限公司 五环三萜类化合物的氨基酸偶合物前药及其医药用途
CN101798333A (zh) * 2009-02-10 2010-08-11 朱永亮 一个三萜化合物及其衍生物用于癌症、炎症及中枢神经系统疾病的治疗
CN101607979A (zh) * 2009-03-02 2009-12-23 中国药科大学 五环三萜-维生素c缀合物、其制备方法及其医药用途

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060166867A1 (en) * 2003-04-28 2006-07-27 Yeda Research And Development Co. Ltd. Novel conjugates of polysaccharides and uses thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HORIUCHI, KUMIKO ET AL.: "Antimicrobial Activity of Oleanolic Acid from Salvia officinalis and Related Compounds on Vancomycin-Resistant Enterococci (VRE", BIOL. PHARM. BULL., vol. 30, no. 6, 16 March 2007 (2007-03-16) *
WANG, CHAO ET AL.: "Conjugation of a Nonspecific Antiviral Sapogenin with a Specific HIV Fusion Inhibitor: A Promising Strategy for Discovering New Antiviral Therapeutics", JOURNAL OF MEDICINAL CHEMISTRY, vol. 57, no. 17, 26 August 2014 (2014-08-26) *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11266632B1 (en) * 2021-07-20 2022-03-08 King Abdulaziz University Maslinic and oleanolic acids derivatives for treating SARS-CoV-2 infection
CN119431488A (zh) * 2024-11-05 2025-02-14 西北大学 齐墩果烷型三氮唑糖苷化合物及其在制备ptp1b抑制剂和抗糖尿病药物中的应用

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