Use of perillyl aldehyde for antibacterial purposes

Abstract

The application discloses a use of a compound with a structure as shown in formula II in preparation of a medicine or reagent for inhibiting or killing drug-resistant or non-drug-resistant bacteria and fungi, wherein R is equal to -CH2OH or -CHO. The application also discloses a use of linalool in preparation of a medicine or reagent for inhibiting or killing drug-resistant or non-drug-resistant bacteria and fungi. Linalool, perilene and perilene aldehyde have antibacterial activity on fungi, various drug-resistant gram-negative bacteria and gram-positive bacteria, and are expected to be developed into new medicines for clinical antibacterial treatment, especially for drug-resistant bacteria.

Description

[0001] This application is a divisional application of the invention entitled "Use of linalool, perillol and perillaldehyde in antibacterial applications" filed on May 6, 2022, with application number 2022104876029. Technical Field

[0002] This invention belongs to the field of natural medicine technology and relates to new uses of linalool, perillol, and perillaldehyde in antibacterial applications. Specifically, it relates to the use of linalool, perillol, and perillaldehyde in the preparation of drugs or reagents for inhibiting or killing extended-spectrum β-lactamase-producing Escherichia coli, methicillin-resistant Staphylococcus aureus, carbapenem-resistant Acinetobacter baumannii, vancomycin-resistant Enterococcus faecalis, carbapenem-resistant Klebsiella pneumoniae, Candida albicans, and Cryptococcus neoformans. It also relates to the use of perillol in the inhibition or killing of carbapenem-resistant Pseudomonas aeruginosa. Background Technology

[0003] The discovery of penicillin and streptomycin is hailed as a major milestone in the history of antibiotic development. The former ushered in the antibiotic era, while the latter opened a new era of large-scale antibiotic screening. The rapid development and application of antibiotics have significantly extended human lifespan, making a vital contribution to human health. With the advancement of science and technology, semi-synthetic antibiotics have emerged, and synthetic antibacterial drugs have been developed, leading to a growing family of antibacterial drugs. Currently discovered mechanisms of antibacterial action mainly include inhibiting the synthesis of bacterial cell walls, proteins, and nucleic acids, as well as inhibiting bacterial folic acid metabolism and cell membrane function.

[0004] With the use of antibiotics, various drug-resistant bacteria have emerged, no longer suppressed by the original antibiotics, posing a significant threat to human health. The WHO has compiled a list of key pathogens for new antibiotic research and development:

[0005] Category 1 (Extremely Important): Carbapenem-resistant Acinetobacter baumannii, carbapenem-resistant Pseudomonas aeruginosa, carbapenem-resistant, extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae.

[0006] Category 2 (extremely important): Vancomycin-resistant Enterococcus faecalis, Methicillin-resistant, Vancomycin-intermediate and drug-resistant Staphylococcus aureus, Clarithromycin-resistant Helicobacter pylori, Fluoroquinolone-resistant Campylobacter, Fluoroquinolone-resistant Salmonella, Cephalosporin-resistant, and Fluoroquinolone-resistant Neisseria gonorrhoeae.

[0007] Category 3 (moderately important): Penicillin-resistant Streptococcus pneumoniae, ampicillin-resistant Haemophilus influenzae, and fluoroquinolone-resistant Shigella spp.

[0008] To address the problem of antibiotic resistance, we need to find new antibacterial drugs.

[0009] Furthermore, in recent years, with the increase in the number of immunocompromised individuals, the widespread use of organ transplants, invasive procedures for the airway or digestive tract, and the extensive routine use of antibiotics, glucocorticoids, and immunosuppressants, the number of patients with invasive fungal infections has increased significantly. Among these, Candida albicans remains the most common pathogen. [1-4] However, antifungal drugs are currently scarce, making the search for such drugs of great significance.

[0010] References

[0011] [1]Pappas Peter G, Kauffman Carol A, Andes David R, et al. ClinicalPractice Guideline for the Management of Candidiasis: 2016 Up date by the Infectious Diseases Society of America. [J]. Clinical infectious diseases: an official publication of the Infectious Diseases Society of America, 2016, 62(4): 409-417.

[0012] [2]Silva-Rocha WP,de Azevedo MF,Chaves GM.Epidemiology and fungalspecies distribution of superficial mycoses in Northeast Bra zil[J].Journalde Mycologie Médicale,2017,27(1):57-64.

[0013] [3]Ibrahim NH,Melake NA,Somily AM,et al.The effect of antifungalcombination on transcripts of a subset of drug-resistance genes in clinicalisolates of Candida species induced biofilms[J].Saudi Pharmaceutical Journal,2015,23(1):55-66.

[0014] [4] Hao Xiaokang, Wang Yuhe. Research progress on the combined effects of drugs on drug-resistant Candida albicans [J]. Medical Theory and Practice, 2021, 34(24):4256-4258. Summary of the Invention

[0015] Perilla (Perilla frutescens (L.) Britt.) is an annual, erect herb belonging to the Lamiaceae family. Its stems, leaves, and fruits are used medicinally. It is cultivated in South Asia, East Asia, and Southeast Asia, and is also widely cultivated in my country. As a plant used for both food and medicine, perilla is rich in various active ingredients and has high medicinal and edible value. The main components of perilla include volatile oils, fatty acids, anthocyanins, flavonoids, triterpenoids, phenolic acids, proteins, and trace elements. The inventors have for the first time discovered that linalool, perillyl alcohol, and perillaldehyde have antibacterial activity against fungi, various drug-resistant Gram-negative bacteria, and Gram-positive bacteria, and hold promise for development into novel antibacterial drugs, especially against drug-resistant bacteria, for clinical use. Furthermore, linalool, perillyl alcohol, and perillaldehyde are found in high concentrations in Lamiaceae plants such as perilla. Perilla has low cultivation costs, and the volatile oil extraction process is simple, significantly reducing the cost of the drug and making it suitable for large-scale production.

[0016] The structures of linalool, perillyl alcohol, and perillaldehyde are shown in Formulas I and II, respectively. a II b As shown:

[0017]

[0018] One of the objectives of this invention is to provide new uses for perillyl alcohol and perillaldehyde in antibacterial applications.

[0019] Use of compounds with structures as described in Formula II in the preparation of drugs or reagents for inhibiting or killing drug-resistant or drug-free bacteria and fungi.

[0020]

[0021] Where R = -CH2OH or -CHO. When R = -CH2OH, the compound is perillyl alcohol; when R = -CHO, the compound is perillaldehyde.

[0022] When the compound is perillyl alcohol, the drug-resistant bacteria are extended-spectrum β-lactamase-producing *Escherichia coli*, carbapenem-resistant *Acinetobacter baumannii*, carbapenem-resistant *Pseudomonas aeruginosa*, methicillin-resistant *Staphylococcus aureus*, vancomycin-resistant *Enterococcus faecium*, and carbapenem-resistant *Klebsiella pneumoniae*. The non-drug-resistant bacteria are *Escherichia coli*, *Acinetobacter baumannii*, *Pseudomonas aeruginosa*, *Staphylococcus aureus*, *Enterococcus faecium*, and *Klebsiella pneumoniae*. The fungi are *Candida albicans* and *Cryptococcus neoformans*.

[0023] When the compound is perillaldehyde, the drug-resistant bacteria are extended-spectrum β-lactamase-producing *Escherichia coli*, carbapenem-resistant *Acinetobacter baumannii*, methicillin-resistant *Staphylococcus aureus*, vancomycin-resistant *Enterococcus faecium*, and carbapenem-resistant *Klebsiella pneumoniae*. The non-drug-resistant bacteria are *Escherichia coli*, *Acinetobacter baumannii*, *Staphylococcus aureus*, *Enterococcus faecium*, and *Klebsiella pneumoniae*. The fungi are *Candida albicans* and *Cryptococcus neoformans*.

[0024] One of the objectives of this invention is to provide new uses for linalool in antibacterial applications.

[0025] Use of linalool in the preparation of drugs or reagents for inhibiting or killing drug-resistant or drug-free bacteria and fungi.

[0026] The drug-resistant bacteria are extended-spectrum β-lactamase-producing *Escherichia coli*, carbapenem-resistant *Acinetobacter baumannii*, methicillin-resistant *Staphylococcus aureus*, vancomycin-resistant *Enterococcus faecium*, and carbapenem-resistant *Klebsiella pneumoniae*. The non-drug-resistant bacteria are *Escherichia coli*, *Acinetobacter baumannii*, *Staphylococcus aureus*, *Enterococcus faecium*, and *Klebsiella pneumoniae*. The fungi are *Candida albicans* and *Cryptococcus neoformans*.

[0027] Another object of the present invention is to provide a pharmaceutical composition for antibacterial purposes, wherein the pharmaceutical composition is a formulation suitable for pharmaceutical use, wherein the main active ingredient is a compound or its derivative having the structure as described in Formula II, and the compound is combined with a pharmaceutically acceptable carrier.

[0028] In the pharmaceutical composition, the compound or its derivative with the structure described in Formula II accounts for 0.1 to 99.9% by weight, and the pharmaceutically acceptable carrier accounts for 0.1 to 99.9% by weight.

[0029] The pharmaceutical composition of the present invention, as a formulation, contains an effective amount of a compound or its derivative with the structure described in Formula II in each dose of 0.1 to 1000 mg. Each dose refers to each formulation unit, such as each tablet or each capsule, or it may refer to the dosage taken at one time, such as 100 mg per dose.

[0030] Another object of the present invention is to provide a pharmaceutical composition for antibacterial purposes, wherein the pharmaceutical composition is formulated into a pharmaceutically acceptable preparation with linalool or its derivatives as the main active ingredient or active ingredient, together with a pharmaceutically acceptable carrier.

[0031] In the pharmaceutical composition, linalool or its derivatives account for 0.1% to 99.9% by weight, and the pharmaceutically acceptable carrier accounts for 0.1% to 99.9% by weight.

[0032] The pharmaceutical composition of the present invention, as a formulation, contains an effective amount of linalool or its derivative in each dose of 0.1 to 1000 mg. Each dose refers to each unit of the formulation, such as each tablet or each capsule, or it may refer to the dosage taken at one time, such as 100 mg per dose.

[0033] The pharmaceutical preparations of this invention include tablets, capsules, flat capsules, granules, pills, powders, ointments, suspensions, injections, powder for injection, suppositories, creams, drops or patches, solutions, suspensions, and emulsions. Specifically, the tablets are sugar-coated tablets, film-coated tablets, enteric-coated tablets, or sustained-release tablets; the capsules are hard capsules, soft capsules, or sustained-release capsules; and the powder for injection is a lyophilized powder for injection.

[0034] When the pharmaceutical compositions of the present invention are prepared into solid or semi-solid pharmaceutical formulations in the form of powder for injection, tablets, powders, capsules, suppositories, and ointments, a solid carrier may be used. The solid carrier may be selected from one or more substances selected from diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, swelling agents, etc., or may be an encapsulating substance. In powder formulations, the carrier contains 5-70% micronized active ingredient. Suitable solid carriers include magnesium carbonate, magnesium stearate, talc, sucrose, lactose, pectin, dextrin, starch, gelatin, methylcellulose, sodium carboxymethyl cellulose, low-boiling-point waxes, cocoa butter, etc. Tablets, powders, suppositories, and capsules represent the most advantageous oral solid dosage forms due to their ease of administration.

[0035] The liquid formulations of this invention include solutions, suspensions, and emulsions. For example, injectable formulations for non-gastrointestinal administration may be in the form of water or water-propylene glycol solutions, with their isotonicity, pH, etc., adjusted to suit physiological conditions in vivo. Liquid formulations may also be prepared as solutions in polyethylene glycol or aqueous solutions. Oral aqueous solutions can be prepared by dissolving the active ingredient in water and then adding appropriate amounts of colorants, flavoring agents, stabilizers, and thickeners. Orally suitable aqueous suspensions can be prepared by dispersing micronized active ingredients in viscous substances such as natural and synthetic gums, methylcellulose, sodium carboxymethyl cellulose, and other known suspending agents.

[0036] For ease of administration and uniform dosage, it is particularly advantageous to formulate the above-mentioned pharmaceutical preparations in unit dosage form. Unit dosage form refers to a physically separated unit suitable for single-dose administration, each unit containing a calculated, predetermined amount of active ingredient to produce the desired therapeutic effect. This unit dosage form can be in package form, such as tablets, capsules, or powders in tubes or vials, or ointments, gels, or creams in tubes or bottles.

[0037] Although the amount of active ingredient contained in the dosage unit can vary, it is generally adjusted to a range of 1 to 1000 mg depending on the potency of the selected active ingredient.

[0038] Those skilled in the art can determine the preferred dosage for a given situation using conventional methods. Generally, the initial treatment dose is lower than the optimal dose of the active ingredient, and the dosage is then gradually increased until the optimal therapeutic effect is achieved. For therapeutic purposes, the total daily dose may be administered once or in divided doses. Detailed Implementation

[0039] The following examples are provided to help those skilled in the art better understand the present invention, but are not intended to limit the invention in any way.

[0040] Example 1

[0041] Determination of the minimum inhibitory concentrations (MICs) of linalool, perillaldehyde, and perillyl alcohol against bacteria and fungi.

[0042] Six types of bacteria: extended-spectrum β-lactamase-producing Escherichia coli (ESBLS), carbapenem-resistant Acinetobacter baumannii (CRAB), carbapenem-resistant Pseudomonas aeruginosa (CRPA), methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecalis (VRE), and carbapenem-resistant Klebsiella pneumoniae (CRE).

[0043] Two fungi: Candida albicans ATCC10231 and Cryptococcus neoformans ATCC208821.

[0044] Procedure: The minimum inhibitory concentrations (MICs) of linalool, perillaldehyde, and perillyl alcohol were determined using the two-fold dilution method in 96-well plates. MH medium was used for bacteria, and SDB medium was used for fungi. The culture volume was 100 μL per well, and the initial bacterial concentration was 5 × 10⁻⁶. 5 The drug concentration in the first well of the drug-treated group was 50 μL / mL (CFU / L). 100 μL of the corresponding culture medium was used as a blank control (MH medium for bacteria, SDB medium for fungi). After incubation at 37℃ for 16 hours, the clarity of the wells was observed, and the wells were labeled. The absorbance at 600 nm was measured using a microplate reader. By comparing the clear wells of the drug-treated group and the blank control group, the minimum inhibitory concentrations (MICs) of linalool, perillaldehyde, and perillyl alcohol against the seven bacteria were calculated. The results are shown in Table 1.

[0045] Table 1. Determination of the minimum inhibitory concentrations (MICs) of linalool, perillaldehyde, and perillol against bacteria and fungi (μL / mL)

[0046] Linalool Perilla alcohol Perillaldehyde Escherichia coli producing extended-spectrum β-lactamase 6.25 1.56 12.5 Carbapenem-resistant Acinetobacter baumannii 1.56 0.0977 6.25 Carbapenem-resistant Pseudomonas aeruginosa ＞50.0 6.25 ＞50.0 Methicillin-resistant Staphylococcus aureus 3.125 0.195 0.195 Vancomycin-resistant Enterococcus faecalis 6.25 0.195 12.5 Carbapenem-resistant Klebsiella pneumoniae 3.13 0.781 12.5 Candida albicans 0.391 ＜0.01 0.391 Cryptococcus neoformans 0.25 ＜0.01 1.5

[0047] Table 1 shows that linalool has inhibitory or bactericidal effects on extended-spectrum β-lactamase-producing Escherichia coli, carbapenem-resistant Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis, carbapenem-resistant Klebsiella pneumoniae, Candida albicans, and Cryptococcus neoformans; perillyl alcohol has inhibitory or bactericidal effects on extended-spectrum β-lactamase-producing Escherichia coli, carbapenem-resistant Acinetobacter baumannii, carbapenem-resistant Pseudomonas aeruginosa, and other bacteria. Methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis, carbapenem-resistant Klebsiella pneumoniae, Candida albicans, and Cryptococcus neoformans have inhibitory or bactericidal effects; perillaldehyde has inhibitory or bactericidal effects on extended-spectrum β-lactamase-producing Escherichia coli, carbapenem-resistant Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis, carbapenem-resistant Klebsiella pneumoniae, Candida albicans, and Cryptococcus neoformans.

[0048] The bacteria and fungi used in the embodiments of this invention are merely illustrative of the fact that "linalool, perillol, and perillaldehyde have inhibitory or bactericidal effects on extended-spectrum β-lactamase-producing Escherichia coli, carbapenem-resistant Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis, carbapenem-resistant Klebsiella pneumoniae, Candida albicans, and Cryptococcus neoformans; and that perillol has inhibitory or bactericidal effects on carbapenem-resistant Pseudomonas aeruginosa." However, the bacteria and fungi used are not limited to these sources. Those skilled in the art should understand that any bacteria or fungi possessing the corresponding bacteria or fungi and identified as producing extended-spectrum β-lactamase-producing Escherichia coli, carbapenem-resistant Acinetobacter baumannii, carbapenem-resistant Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis, carbapenem-resistant Klebsiella pneumoniae, Candida albicans, or Cryptococcus neoformans are applicable to this invention.

Claims

1. The use of the compound with the structure of Formula II in the preparation of drugs or reagents for inhibiting or killing drug-resistant bacteria and fungi; wherein the drug-resistant bacteria are extended-spectrum β-lactamase-producing Escherichia coli, carbapenem-resistant Acinetobacter baumannii, vancomycin-resistant Enterococcus faecalis, and carbapenem-resistant Klebsiella pneumoniae; and wherein the fungus is Cryptococcus neoformans. ； in, R=-CHO.

2. The use according to claim 1, characterized in that: The drug is a formulation made from a compound with the structure described in Formula II as the active ingredient and a drug-acceptable carrier.

Images