Polyaminated fatty acids for use as potentiators of antibacterial activity

Abstract

The present invention relates to the use of a polyaminated fatty acid compound corresponding to formula (I), the pharmaceutically acceptable salts thereof and / or the solvates thereof as adjuvants for antibacterial agents, a pharmaceutical composition comprising at least said polyaminated fatty acid compound and at least one antibacterial agent, said pharmaceutical composition being used as a medicament, in particular as a medicament. The pharmaceutical composition is used for treating bacterial diseases or infectious diseases, especially bacterial diseases or infectious diseases caused by Gram-negative bacteria, and is used for killing holding bacteria cells or inhibiting the growth of the holding bacteria cells. A kit comprising a composition comprising said polyaminated fatty acid compound and a separate pharmaceutical composition comprising at least one antibacterial agent, as well as a polyaminated fatty acid compound corresponding to formula (I '-a), a pharmaceutically acceptable salt thereof and / or a solvate thereof.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480024204.0 (22) Application Date 2024.04.05 (30) Priority Data 23305524.3 2023.04.07 EP (85) PCT International Application Entering National Phase Date 2025.09.30 (86) PCT International Application Application Data PCT / EP2024 / 059350 2024.04.05 (87) PCT International Application Publication Data WO2024 / 209060 EN 2024.10.10 (71) Applicant Aix-Marseille University Address Marseille, France Applicant National Institute of Health French National Centre for Scientific Research (72) Inventor Jean-Michel Brunel (74) Patent Agency Beijing Baishansong Intellectual Property Agency (General Partnership) 11413 Patent Attorneys Li Shengnan and Wang Chunwei (51) Int.Cl. A61K 31 / 16 (2006.01) A61L 2 / 00 (2006.01) C07C 233 / 38 (2006.01) A61P 31 / 04 (2006.01) (54) Invention Title: Polyamined Fatty Acids Used as Enhancers of Antibacterial Activity (57) Abstract: This invention relates to the use of polyamined fatty acid compounds corresponding to formula (I), pharmaceutically acceptable salts thereof, and / or solvates thereof as adjuvants to antibacterial drugs; a pharmaceutical composition comprising at least the polyamined fatty acid compound and at least one antibacterial drug, the pharmaceutical composition being used as a drug for treating bacterial diseases or infectious diseases, particularly bacterial diseases or infectious diseases caused by Gram-negative bacteria, the pharmaceutical composition being used to kill resident bacterial cells or inhibit the growth of resident bacterial cells; a kit comprising a composition comprising the polyamined fatty acid compound and a separate pharmaceutical composition comprising at least one antibacterial drug; and a polyamined fatty acid compound corresponding to formula (I'-a), pharmaceutically acceptable salts thereof, and / or solvates thereof. Claims 8 pages, Description 33 pages, Drawings 2 pages, CN 121443283 A 2026.01.30 CN 1 21 44 32 83 A 1. Use of a polyamined fatty acid compound corresponding to the following formula (I), its pharmaceutically acceptable salt and / or its solvate as an adjuvant to an antibacterial agent: R1-C(=O)-NH-(CR2R3)m-[X-(CR 4R5)n]p-NR 6R7 (I) Wherein: -R1 represents a straight-chain aliphatic chain containing at least 7 carbon atoms,The straight-chain aliphatic chain is a monounsaturated aliphatic chain, a polyunsaturated aliphatic chain, or a saturated aliphatic chain; -R2 and R3 independently represent a hydrogen atom or a C1-C8 alkyl group in each of their m occurrences; -R4 and R5 independently represent a hydrogen atom or a C1-C8 alkyl group in each of their n occurrences and p occurrences; -X independently represents an -NR8- group or a divalent 5- to 7-membered heterocyclic alkyl group containing at least one nitrogen atom in each of its p occurrences; wherein R8 represents a hydrogen atom, a C1-C6 alkyl group, or -(CH2)q-NH2, where q represents an integer from 1 to 5; -R6 and R7 independently represent hydrogen atoms, C1-C8 alkyl groups, or R6 and R7 together with the nitrogen atoms to which they are attached form 5- to 7-membered heterocyclic groups optionally substituted with one to three R9s; wherein R9 represents -(CH2)u-NH2, where u represents an integer from 1 to 5; -m is an integer from 2 to 10; -n independently represents an integer from 1 to 5 in each of its p occurrences; and -p is an integer from 0 to 4. 2. The use according to claim 1, wherein the straight-chain aliphatic chain R1 is a monounsaturated aliphatic chain or a polyunsaturated aliphatic chain. 3. The use according to claim 1 or claim 2, wherein R2 and R3 independently represent hydrogen atoms or C1-C3 alkyl groups in each of their m occurrences. 4. The use according to any of the preceding claims, wherein R4 and R5 independently represent hydrogen atoms or C1-C3 alkyl groups in each of their n occurrences and p occurrences. 5. The use according to any one of the preceding claims, wherein p is 1 to 3. 6. The use according to any one of the preceding claims, wherein X independently represents a -NR8- group, a C1-C3 alkyl group, or -(CH2)q-NH2 in each of the p occurrences, wherein R8 represents a hydrogen atom, and wherein q represents an integer from 1 to 3. 7. The use according to any one of the preceding claims, wherein R6 and R7 are hydrogen atoms. 8. The use according to any one of the preceding claims, wherein the polyamined fatty acid compound is selected from the following compounds, their pharmaceutically acceptable salts, and / or their solvates: Claims 1 / 8 page 2 CN 121443283 A Claims 2 / 8 page 3 CN 121443283 A Claims 3 / 8 page 4 CN 121443283 A Claims 4 / 8 page 5 CN 121443283 A 9. A pharmaceutical composition comprising at least one polyamined fatty acid compound as defined in any one of the preceding claims and at least one antibacterial agent. 10. The pharmaceutical composition according to claim 9,Based on the total weight of the pharmaceutical composition, the pharmaceutical composition comprises 0.1% to 50% by weight of the polyamined fatty acid compound. 11. The pharmaceutical composition according to claim 9 or claim 10, wherein the weight ratio of the polyamined fatty acid compound to the antibacterial agent is 0.2 to 2. 12. The pharmaceutical composition according to any one of claims 9 to 11, wherein the antibacterial agent is selected from tetracyclines and macrolides. 13. A pharmaceutical composition according to any one of claims 9 to 12, used as a medicine. 14. A pharmaceutical composition for use according to claim 13, used to treat bacterial diseases; particularly bacterial diseases caused by Gram-negative bacteria. 15. A pharmaceutical composition for use according to claim 13, used to kill resident bacterial cells or inhibit the growth of resident bacterial cells. 16. A kit comprising: - a pharmaceutical composition comprising the polyamined fatty acid compound as defined in any one of claims 1 to 8, and - a separate pharmaceutical composition comprising at least one antibacterial agent. 17. A polyamined fatty acid compound selected from compounds of the following formula (I'-a): Claims 5 / 8 page 6 CN 121443283 A Claims 6 / 8 page 7 CN 121443283 A Claims 7 / 8 page 8 CN 121443283 A Pharmaceutically acceptable salts and / or solvates thereof. Claims 8 / 8 Page 9 CN 121443283 A Polyamined fatty acids used as enhancers of antibacterial activity

[0001] The present invention relates to the use of polyamined fatty acid compounds corresponding to formula (I), pharmaceutically acceptable salts thereof and / or solvates thereof as adjuvants to antibacterial drugs, a pharmaceutical composition comprising at least the polyamined fatty acid compound and at least one antibacterial drug, the pharmaceutical composition being used as a drug for treating bacterial diseases or infectious diseases, particularly bacterial diseases or infectious diseases caused by Gram-negative bacteria, the pharmaceutical composition being used to kill or inhibit the growth of persistent bacterial cells, a kit comprising a composition containing the polyamined fatty acid compound and a separate pharmaceutical composition containing at least one antibacterial drug, and a specific polyamined fatty acid compound corresponding to formula (I'-a), pharmaceutically acceptable salts thereof and / or solvates thereof.

[0002] The number of immunocompromised populations at serious risk of infection is increasing. These groups include people over 65 years of age, newborns, people with diabetes, chemotherapy patients, and surgical patients, both in hospitals and in the community worldwide. These groups are most likely to contract severe infections. The most common hospital-acquired infections are pneumonia and urinary tract infections (UTIs).It can be caused by a variety of different bacterial species. Pneumoniae infection accounts for 15% of all hospital-acquired infections and 24% of intensive care unit-acquired infections. Among the most common hospital-acquired infections, severe pneumonia has the highest mortality rate, is the sixth leading cause of death, and is also a leading cause of sepsis.

[0003] The current situation of serious hospital-acquired infections is worrying because some infections are no longer treatable with existing drugs due to the emergence of new pathogens with enhanced and difficult-to-overcome antibiotic resistance. In fact, in human and animal health, whether in cities or hospitals, the large-scale and repeated use of antibiotics, and their misuse (e.g., treatment for too short, too long, or inappropriate dosage), has led to the emergence of bacterial strains resistant to these antibiotics. Resistance was initially sporadic but has now become widespread and worrying. Multidrug resistance (MDR) is a phenomenon attributed to various mechanisms expressed in the same bacterial strain against different families of antibiotics. In February 2017, the WHO published a list of antibiotic-resistant "priority pathogens" that pose the greatest threat to human health. The most critical group of pathogens includes multidrug-resistant bacteria, which pose a particular threat in hospitals, nursing homes, and among patients requiring care with equipment such as ventilators and blood catheters. These bacteria include *Acinetobacter*, *Pseudomonas*, and various *Enterobacteriaceae*, including *Klebsiella*, *Escherichia coli*, *Serratia*, and *Proteus*. They can cause serious and often fatal infections, such as bloodstream infections and pneumonia.

[0004] These drug-resistant strains have become particularly prevalent and deadly in Europe, the United States, Africa, and Asia, leading to high morbidity and mortality rates from hospital-acquired infections (HAIs). Current hospital guidelines generally recommend combination antibiotic therapy for pneumonia in order to treat these hospital-acquired infections and improve the likelihood of adequate antibiotic coverage. While using more than one antibiotic to treat infections can effectively target many different bacteria, these guidelines only provide a temporary solution to the problem of antibiotic resistance and will ultimately lead to more harm, as increased antibiotic use will only contribute to the evolution of broader antibiotic resistance.

[0005] In addition to hospital-acquired infections and antibiotic resistance, particularly with Gram-negative bacteria, there is a daunting and critical unmet medical need as antibiotic development and federal approval are declining. Therefore, new treatment strategies are needed to improve antibiotic efficacy and / or reduce resistance mechanisms. National and international health agencies have taken a series of measures,This is to encourage controlled drug use and the development of new molecules specifically for treating Gram-negative bacteria. The low permeability of these bacteria—surrounded by an outer membrane that reduces compound diffusion—and their constitutive expression of efflux pumps (protein complexes, CN 121443283 A, which actively transport toxic molecules out of the cell) result in a limited number of new active molecules. Furthermore, there is ample evidence that they can overproduce these efflux pumps in response to extracellular compounds, including drugs. Historically, improvements in antibiotic activity have largely been achieved by altering the design of pioneer bioactive molecules. New classes of antibiotics have emerged since the 1980s, although they are primarily active against Gram-positive bacteria. Recent target-based high-throughput screening projects, along with computer simulation studies, have led to the identification of high-potential hits. While this strategy appears attractive, a major drawback of target-based assays is that they do not account for membrane translocation barriers, including bacterial permeability and efflux pump issues.

[0006] For many years, researchers have been investigating the use of one or more compounds lacking antimicrobial activity to protect and enhance the effects of existing antibiotics. These special compounds are called "adjuvants." The concept of antibiotic adjuvants involves a molecular enhancement and improvement of the effect of antibiotics against resistant microorganisms. Also known as "resistance circuit breaker," "chemosensitizer," or "antibiotic potentiator," adjuvants are compounds that lack antimicrobial activity, although they may exert a slight inhibitory effect on bacterial growth. In short, they can be thought of as "inhibiting" resistance and "improving" the inhibitory effect when co-administered with an antibiotic.

[0007] For example, International Application WO2021 / 211123A1 describes tryptamine ureas and its derivatives as adjuvants to sensitize Gram-negative bacteria to the action of polymyxin antibiotics such as colistin. Combination therapy with polymyxin antibiotics and adjuvants has proven practical in treating Gram-negative bacterial infections, including those caused by drug-resistant strains.

[0008] Furthermore, drug resistance is not the only obstacle to the efficacy of existing antibiotics. In fact, even high doses of antibiotics may fail to eliminate "dormant" cells, particularly persistent bacteria and viable but non-culturable cells (VBNCs). Surviving dormant cells play a crucial role in infection recurrence. In addition to limiting the effectiveness of antimicrobial agents,Dormant cells can be a cause of chronic diseases. The persistence and activation of dormant cells are closely related to individual and environmental factors, compared to drug resistance.

[0009] Currently, there are very few options available for eliminating dormant cells because most treatments are designed to kill active cells. One strategy specifically targeting persistent bacteria is to use drugs, such as antimicrobial peptides, which interact directly with the cell membrane and are therefore effective even against metabolically inactive bacteria. Another option is to first reactivate dormant cells, for example with sugars, and then use common antimicrobial agents to eliminate the reactivated cells. However, the applicability of these methods is limited; and to date, there is no convenient therapeutic solution for removing persistent bacteria.

[0010] Therefore, there remains a need for new compounds that can restore or enhance the antimicrobial activity of antibiotics, more particularly, that can restore or enhance the antimicrobial activity of existing antibiotics for treating Gram-negative resistant strains, and / or that can induce an anti-persistent bacterial effect of antibiotics while ensuring low toxicity to humans and animals, and whose structure can be readily modified to suit the type of bacteria.

[0011] Through in-depth research, the inventors discovered that polyamined fatty acids can enhance or restore the antibacterial activity of antibiotics against drug-resistant bacteria. They found that these polyamined fatty acids can significantly enhance the antibacterial activity of tetracycline-based antibacterial drugs against drug-resistant bacteria.

[0012] Therapeutic Applications

[0013] A first objective of the present invention is the use of polyamined fatty acid compounds corresponding to the following formula (I), pharmaceutically acceptable salts thereof, and / or solvates thereof, as adjuvants to antibacterial agents:

[0014] R1-C(=O)-NH-(CR2R3)m-[X-(CR4R5)n]p-NR6R7 (I)

[0015] Wherein: Specification 2 / 33 pages 11 CN 121443283 A

[0016] -R1 represents a straight-chain aliphatic chain containing at least 7 carbon atoms, wherein the straight-chain aliphatic chain is a monounsaturated aliphatic chain or a polyunsaturated aliphatic chain, or a saturated aliphatic chain;

[0017] -R2 and R3 independently represent hydrogen atoms or C1-C8 alkyl groups in each of their m occurrences;

[0018] -R4 and R5 independently represent a hydrogen atom or a C1-C8 alkyl group in each of their nth occurrences and in each of their pth occurrences;

[0019] -X independently represents a -NR8- group or a divalent 5- to 7-membered heterocyclic alkyl group containing at least one nitrogen atom in each of its pth occurrences; wherein R8 represents a hydrogen atom, a C1-C6 alkyl group, or -(CH2)q-NH2, where q represents an integer from 1 to 5;

[0020] -R6 and R7 independently represent a hydrogen atom, a C1-C8 alkyl group,Alternatively, R6 and R7 together with the nitrogen atom to which they are attached form a 5- to 7-membered heterocyclic group optionally substituted by one to three R9s; wherein R9 represents -(CH2)u-NH2, where u represents an integer from 1 to 5;

[0021] -m is an integer from 2 to 10;

[0022] -n independently represents an integer from 1 to 5 in each of its p occurrences; and

[0023] -p is an integer from 0 to 4.

[0024] Surprisingly, polyamined fatty acid compounds as defined in this invention are antibiotic enhancers and exhibit a powerful effect on antibiotic sensitivity levels against drug-resistant bacterial strains, and more particularly against Gram-negative drug-resistant bacterial strains.

[0025] Furthermore, such polyamined fatty acid compounds do not exhibit intrinsic antibacterial activity, so that they do not induce resistance mechanisms, although they can greatly restore or enhance antibacterial activity. They may act through membrane depolarization and / or integrity membrane disruption, making their mechanism of action significantly reduce the development of resistance.

[0026] In this invention, the term "antimicrobial agent" refers to a substance or compound that kills or inhibits bacterial growth. Antimicrobial agents are typically administered in cases of bacterial diseases or infections.

[0027] In this invention, the term "polyamined fatty acid compound" includes polyamined fatty acid compounds of formula (I), as well as pharmaceutically acceptable salts and / or solvates thereof.

[0028] Aliphatic chain R1

[0029] R1 preferably contains at least 9 carbon atoms, more preferably at least 11 carbon atoms, and even more preferably at least 13 carbon atoms.

[0030] R1 preferably contains up to 35 carbon atoms, more preferably up to 27 carbon atoms, and even more preferably up to 23 carbon atoms.

[0031] R1 preferably contains 9 to 35 carbon atoms, more preferably 11 to 27 carbon atoms, and even more preferably 13 to 23 carbon atoms.

[0032] The straight-chain aliphatic chain R1 is a monounsaturated aliphatic chain or a polyunsaturated aliphatic chain, or a saturated aliphatic chain, and preferably a monounsaturated aliphatic chain or a polyunsaturated aliphatic chain. Monounsaturated aliphatic chains or polyunsaturated aliphatic chains result in enhanced antibacterial activity.

[0033] The monounsaturated straight-chain aliphatic chain or polyunsaturated straight-chain aliphatic chain R1 may contain 1 to 6 unsaturated bonds, preferably 2 to 5 unsaturated bonds, and more preferably 2 to 4 unsaturated bonds. The latter two ranges result in lower toxicity to mammals.

[0034] Each unsaturated bond may be Z-configuration (cis configuration) or E-configuration (trans configuration). In other words,The polyamined fatty acid compound of this invention, page 3 / 33, CN 121443283 A, can exist in the form of a mixture of isomers or in the form of a single isomer. Preferably, the polyamined fatty acid compound of this invention exists in the form of a single isomer.

[0035] In this invention, the term "aliphatic chain" refers to a hydrocarbon chain, and the term "aliphatic" is the opposite of "aromatic".

[0036] In this invention, the term "straight chain" refers to an unsubstituted or unbranched aliphatic chain.

[0037] The linear aliphatic chain R1 is preferably selected from lauric acid, stearic acid, caproic acid, myristic acid, palmitic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid, montanic acid, melissic acid, trichosanoic acid, oleic acid, linoleic acid, lauroleic acid, myristoleic acid, palmitoleic acid, trans-octadecanoic acid, gadoleic acid, ketoleic acid, erucic acid, selacholeic acid (nervonic acid), linolenic acid, stearidonic acid, eleostearic acid, arachidonic acid, cuplanodonic acid, and elaidic acid. Fatty acid chains of docosahexanoic acid, docosahexaenoic acid, docosapentaenoic acid, docosapentaenoic acid, eicosapentaenoic acid, and petroselinic acid.

[0038] In a preferred embodiment,The straight-chain aliphatic chain R1 is preferably a fatty acid chain selected from lauric acid, stearic acid, palmitic acid, behenic acid, lignoceric acid, trisaccharide acid, oleic acid, linoleic acid, myristoleic acid, palmitic acid, squalane, arachidonic acid, and docasahexanoic acid.

[0039] R2 and R3

[0040] R2 and R3 independently represent a hydrogen atom or a C1-C8 alkyl group in each of their m occurrences.

[0041] The C1-C8 alkyl group can be a straight-chain or branched alkyl group, preferably a straight-chain alkyl group or an alkyl group with branches containing 1 to 3 carbon atoms, and more preferably a straight-chain alkyl group.

[0042] In a preferred embodiment, R2 and R3 independently represent a hydrogen atom or a C1-C5 alkyl group in each of their m occurrences, and more preferably a hydrogen atom or a C1-C3 alkyl group.

[0043] Advantageously, at least one of R2 and R3 is a hydrogen atom, and more advantageously both R2 and R3 are hydrogen atoms.

[0044] m represents the number of times the -CR2R3- group appears. m is 2 to 10, preferably 2 to 6, and more preferably 2 to 4.

[0045] R2 and R3 are chosen independently in each occurrence of m occurrences. In other words, this means that for each occurrence of m occurrences, the (R2, R3) pair (or -CR2R3- group) can be the same or different. In fact, R2 and R3 can take different meanings in each occurrence of m occurrences.

[0046] In a preferred embodiment, the (R2, R3) pair (or -CR2R3- group) is the same in each occurrence of m occurrences.

[0047] R4 and R5

[0048] R4 and R5 independently represent hydrogen atoms or C1-C8 alkyl groups in each occurrence of n occurrences and in each occurrence of p occurrences.

[0049] The C1-C8 alkyl group can be a straight-chain or branched alkyl group, preferably a straight-chain alkyl group or an alkyl group with branches containing 1 to 3 carbon atoms, and more preferably a straight-chain alkyl group.

[0050] In a preferred embodiment, R4 and R5 independently represent hydrogen atoms or C1-C5 alkyl groups, and more preferably hydrogen atoms or C1-C3 alkyl groups, in each occurrence of n occurrences and in each occurrence of p occurrences.

[0051] Advantageously, at least one of R4 and R5 is a hydrogen atom, and more advantageously both R4 and R5 are hydrogen atoms. Specification 4 / 33 pages 13 CN 121443283 A

[0052] n represents the number of times the -CR4R5- group appears. n is 1 to 5, preferably 2 to 5, and more preferably 2 to 4.

[0053] R4 and R5 are independently selected in each occurrence of n occurrences. In other words, this means that when n is greater than 1,The (R4, R5) pair (or -CR4R5- group) can be the same or different in each of the n occurrences. In fact, R4 and R5 can take different meanings in each of the n occurrences.

[0054] In a preferred embodiment, the (R4, R5) pair (or -CR4R5- group) is the same in each of the n occurrences.

[0055] p represents the number of times the [X-(CR4R5)n] group appears.

[0056] p is 0 to 4, and preferably 1 to 3, and more preferably 2 to 3.

[0057] R4 and R5 are chosen independently in each of the p occurrences. In other words, this means that when p is greater than 1, the (R4, R5) pair (or -CR4R5- group) can be the same or different in each of the p occurrences. In fact, R4 and R5 can take different meanings in each of the p occurrences.

[0058] Furthermore, n independently represents an integer from 1 to 5 in each of the p occurrences. In other words, when p is greater than 1, n can take different values ​​or meanings in each of the p occurrences.

[0059] Advantageously, the number of occurrences n is equal to 4 at least once.

[0060] X

[0061] X independently represents a -NR8- group or a divalent 5- to 7-membered heterocyclic alkyl group containing at least one nitrogen atom in each of the p occurrences; wherein R8 represents a hydrogen atom, a C1-C6 alkyl group, or -(CH2)q-NH2, wherein q represents an integer from 1 to 5.

[0062] X is independently selected in each of the p occurrences. In other words, this means that X can be the same or different for each of the p occurrences. In fact, X can take different meanings in each of the p occurrences.

[0063] In this invention, the term "heterocyclic alkyl" refers to a cyclic alkyl group containing 2 to 7 carbon atoms and one or more heteroatoms, wherein the one or more heteroatoms are preferably selected from oxygen atoms, nitrogen atoms, and mixtures thereof.

[0064] A divalent 5- to 7-membered heterocyclic alkyl group (as group X) containing at least one nitrogen atom preferably contains 4 to 6 carbon atoms and at least one nitrogen atom as a heteroatom. In a preferred embodiment, the heterocyclic alkyl group is 5- or 6-membered, and more preferably 6-membered. The heterocyclic alkyl group preferably contains one or two nitrogen atoms as heteroatoms.

[0065] For R8, a hydrogen atom or a -(CH2)q-NH2 group is preferred.

[0066] The C1-C6 alkyl group can be a straight-chain or branched alkyl group, preferably a straight-chain alkyl group or an alkyl group with branches containing 1 to 3 carbon atoms, and more preferably a straight-chain alkyl group.

[0067] In a preferred embodiment, X independently represents a -NR8- group or a divalent 5- to 7-membered heterocyclic alkyl group containing at least one nitrogen atom in each of its p occurrences; wherein R8 represents a hydrogen atom, a C1-C3 alkyl group, or -(CH2)q-NH2,Wherein q represents an integer from 1 to 3, and more preferably -NR8- group, wherein R8 represents a hydrogen atom, C1-C3 alkyl, or -(CH2)q-NH2, wherein q represents an integer from 1 to 3.

[0068] Examples of divalent 5- to 7-membered heterocyclic alkyl groups (which are X groups) containing at least one nitrogen atom are piperidine and piperazine.

[0069] R6 and R7

[0070] R6 and R7 independently represent hydrogen atoms, C1-C8 alkyl, or R6 and R7 together with the nitrogen atoms to which they are attached form 5- to 7-membered heterocyclic groups optionally substituted with one to three R9s; wherein R9 represents -(CH2)u-NH2, wherein u represents an integer from 1 to 5.

[0071] In this invention, the term "heterocyclic group" refers to "heterocyclic alkyl" and "heteroaryl" groups as defined above. Specification 5 / 33 pages 14 CN 121443283 A

[0072] In this invention, the term "heteroaryl" refers to an aromatic ring or aromatic ring system containing 5 to 12 carbon atoms, preferably 5 to 10 carbon atoms, having one or two rings fused together or covalently linked, wherein at least one ring is aromatic, and one or more of these rings has one or more carbon atoms substituted by one or more heteroatoms, said one or more heteroatoms preferably selected from oxygen atoms, nitrogen atoms and mixtures thereof.

[0073] C1-C8 alkyl can be straight-chain or branched alkyl, preferably straight-chain alkyl or alkyl with branches containing 1 to 3 carbon atoms, and more preferably straight-chain alkyl.

[0074] In a preferred embodiment, R6 and R7 independently represent hydrogen atoms, C1-C5 alkyl groups, or R6 and R7 together with the nitrogen atoms to which they are attached form 5- to 7-membered heterocyclic groups optionally substituted by one to three R9s; wherein R9 represents -(CH2)u-NH2, where u represents an integer from 1 to 5, and more preferably hydrogen atoms, C1-C3 alkyl groups, or R6 and R7 together with the nitrogen atoms to which they are attached form 5- to 7-membered heterocyclic groups optionally substituted by one to three R9s; wherein R9 represents -(CH2)u-NH2, where u represents an integer from 1 to 5.

[0075] u preferably represents an integer from 2 to 4.

[0076] The 5- to 7-membered heterocyclic groups are preferably unsubstituted or substituted by one R9.

[0077] The 5- to 7-membered heterocyclic groups are preferably selected from piperazine and imidazolyl groups.

[0078] Examples of 5- to 7-membered heterocyclic groups formed by R6, R7, and the nitrogen atom to which they are attached, optionally substituted with one to three R9s, are as follows:

[0079]

[0080] Advantageously, at least one of R6 and R7 is a hydrogen atom,And more advantageously, both R6 and R7 are hydrogen atoms.

[0081] -NH-(CR2R3)m-[X-(CR 4R5)n]p-NR 6R7

[0082] The group -NH-(CR2R3)m-[X-(CR 4R5)n]p-NR 6R7 is preferably selected from the following groups:

[0083]

[0084] In a particularly preferred embodiment, the polyamined fatty acid compound is selected from the following polyamined fatty acid compounds:

[0085]

[0086]

[0087]

[0088]

[0089] pharmaceutically acceptable salts and / or solvates thereof.

[0090] Among the above-described polyamined fatty acid compounds, those having a polyunsaturated aliphatic chain as an R1 group are particularly preferred.

[0091] Polyamined fatty acid compounds are used as adjuvants to antibacterial drugs. As described in the background art, adjuvants are also called "resistance loop blockers," "chemical sensitizers," or "(antibiotic) enhancers."

[0092] Antibacterial drugs and polyamined fatty acid compounds are applied together in an amount that inhibits bacteria more effectively than the same amount of antibacterial drug applied without the application of the polyamined fatty acid compound.

[0093] Preferably, if an enhancing or restorative effect is observed when the polyamined fatty acid compound is applied at a concentration Ca (in μM), where Ca ≤ MICa / 4, and MICa represents the minimum inhibitory concentration (in μM) of the polyamined fatty acid compound against a given bacterial strain, then the polyamined fatty acid compound can be defined as an adjuvant to the antibacterial drug. Instructions for Use, Page 9 / 33, 18 CN 121443283 A

[0094] When the MIC ratio is greater than 1, wherein the MIC ratio = MIC (in μM) of the antimicrobial agent alone against the strain / MIC (in μM) of the antimicrobial agent in the presence of the compound, an enhancing or restorative effect is observed.

[0095] The adjuvant and the antimicrobial agent may be administered simultaneously, separately, or sequentially.

[0096] In the use of the present invention,Antibiotics are preferably selected from tetracyclines and macrolides.

[0097] Tetracyclines are particularly preferred.

[0098] Examples of tetracyclines are tetracycline, doxycycline, minocycline, or tigecycline.

[0099] Examples of macrolides are erythromycin, clarithromycin, azithromycin, josamycin, roxithromycin, spiramycin, tererythromycin, or tylosin.

[0100] Pharmaceutically acceptable salts include acetates, adipic acid salts, aspartate salts, benzoates, benzenesulfonates, bicarbonates / carbonates, bisulfates / sulfates, borates, camphor sulfonates, citrates, cyclosulfonates, ethanedisulfonates, ethanesulfonates, formates, fumarates, gluconate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromine, hydroiodide / iodide, hydroxyethanesulfonate, lactate, malate, maleate, malonate, methanesulfonate, methyl sulfate, naphthylate, 2-napsylate, nicotinate, nitrates, orotate, oxalate, palmitate, papoate, phosphates / hydrogen phosphates / dihydrogen phosphates, pyroglutamate, sucrose, stearates, succinates, tannates, and tartrates. Toluenesulfonate, trifluoroacetate, and xinafoate.

[0101] Pharmaceutically acceptable salts of the compound (I) of the present invention can be prepared by reacting the compound (I) with a suitable acid or by using a suitable ion exchange column. All these reactions are generally carried out in solution. The salt can be precipitated from the solution and collected by filtration, or it can be recovered by evaporating the solvent. The degree of ionization in the salt can vary from complete ionization to almost no ionization.

[0102] A “solvate” refers to a molecular complex comprising one or more molecules of the compound (I) and one or more solvents in stoichiometric or substoichiometric amounts. Typically, the solvent is a pharmaceutically acceptable solvent, such as ethanol. The term “hydrate” refers to a solvate when the solvent is water (H2O). Solvates of the compound (I) include conventional solvates, such as those formed in the final steps of the preparation of these compounds due to the presence of a solvent.

[0103] Polyamined fatty acid compounds as defined in the first objective of this invention are particularly useful as adjuvants to antibacterial drugs for the treatment of bacterial diseases or infections, especially those caused by Gram-negative bacteria.

[0104] Therefore, polyamined fatty acid compounds as defined in the first objective of this invention can be used to treat bacterial diseases or infections, especially those caused by Gram-negative bacteria.

[0105] This invention also relates to polyamined fatty acid compounds as defined in the first objective of this invention.It is used for medical purposes.

[0106] “Gram-negative bacteria” refers to bacteria that do not retain crystal violet staining when Gram staining is used. Gram-negative bacteria are characterized by a bacterial cell wall composed of a thin layer of peptidoglycan, which lies between the inner cytoplasmic cell membrane and the outer bacterial membrane.

[0107] Gram-negative bacteria are preferably selected from Pseudomonas, Escherichia, Klebsiella, Acinetobacter, Enterobacter and Legionella bacteria, and more preferably from Pseudomonas and Escherichia.

[0108] In a preferred embodiment, the Pseudomonas bacteria is Pseudomonas aeruginosa. Instructions for Use, Page 10 / 33, 19 CN 121443283 A

[0109] In a preferred embodiment, the *Escherichia* bacteria is *Escherichia coli*.

[0110] In a preferred embodiment, the *Klebsiella* bacteria is *Klebsiella pneumoniae*.

[0111] In a preferred embodiment, the *Acinetobacter* bacteria is *Acinetobacter baumannii*.

[0112] Polyamined fatty acid compounds as defined in the first objective of the present invention are also particularly useful as adjuvants to antibacterial drugs for killing or inhibiting the growth of resident bacterial cells.

[0113] Polyamined fatty acid compounds as defined in the first objective of the present invention can be prepared by reacting natural fatty acids with polyamines in the presence of a coupling agent such as BOP, a base such as diisopropylethylamine, and in an organic solvent such as dichloromethane. This reaction is typically carried out at room temperature. The following schematic diagram illustrates the reaction.

[0114]

[0115] A second objective of the present invention is a pharmaceutical composition comprising at least one polyamined fatty acid compound as defined in the first objective of the present invention.

[0116] Preferably, the pharmaceutical composition comprises at least one polyamined fatty acid compound as defined in the first objective of the present invention and at least one antibacterial agent.

[0117] In fact, the combination of the polyamined fatty acid compound as defined in the first objective of the present invention with the antibacterial agent produces a pharmaceutical composition having improved antibacterial activity, particularly improved antibacterial activity against antibacterial resistant bacteria, and / or having anti-persistence activity against dormant cells.

[0118] In a preferred embodiment, based on the total weight of the pharmaceutical composition,The pharmaceutical composition comprises about 0.1% by weight to about 50% by weight of the polyamined fatty acid compound, and more preferably about 0.2% by weight to about 30% by weight of the polyamined fatty acid compound.

[0119] The pharmaceutical composition preferably comprises a therapeutically effective amount of an antibacterial agent.

[0120] A “therapeutically effective amount” (hereinafter referred to as “effective amount”) means an amount of therapeutic agent sufficient to achieve a desired therapeutic, preventive, or prophylactic effect in the subject to which it is administered without causing significant negative or adverse side effects to the patient. A therapeutically effective amount may be administered before the onset of a disease, condition, or symptom to prevent disease or preventive action. Alternatively, or additionally, a therapeutically effective amount may be administered after the onset of a disease, condition, or symptom to treat.

[0121] In a preferred embodiment, based on the total weight of the pharmaceutical composition, the pharmaceutical composition comprises about 50% by weight to about 99.9% by weight of the antibacterial agent, and more preferably about 70% by weight to about 99.8% by weight of the antibacterial agent.

[0122] In the pharmaceutical composition, the weight ratio of the polyamined fatty acid compound to the antibacterial agent is preferably from about 0.2 to about 2, and more preferably from about 0.5 to about 1.

[0123] The antibacterial agent is as defined in the first objective of the present invention.

[0124] The pharmaceutical composition may also contain a pharmaceutically acceptable carrier.

[0125] "Pharmaceutically acceptable" means that the components of the pharmaceutical composition are compatible with each other and harmless to the patient to whom it is administered.

[0126] "Pharmaceutically acceptable carrier" means an excipient that does not produce harmful, allergic or other adverse reactions when administered to animals or humans. It includes all solvents, dispersion media, coatings, antifungal agents, isotonic and absorption delay agents, etc. For human administration, the formulation shall meet the standards of sterility, pyrogen-free, general safety and purity as required by regulatory agencies (e.g., the FDA office or EMA).

[0127] Examples of pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances (e.g., sodium carboxymethyl cellulose), polyethylene glycol, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin.

[0128] A third objective of the present invention is a pharmaceutical composition as defined in the second objective of the present invention, which is used as a medicine or for medical use.

[0129] A fourth objective of the present invention is a pharmaceutical composition as defined in the second objective of the present invention.It is used to treat bacterial diseases; particularly bacterial diseases caused by Gram-negative bacteria.

[0130] The bacterial diseases may be selected from chronic infections, recurrent infections, refractory infections, persistent infections, persistent bacteria-associated infections, persistent bacteria-associated chronic infections, persistent bacteria-associated recurrent infections, persistent bacteria-associated refractory infections, and persistent bacteria-associated persistent infections.

[0131] A fifth objective of the present invention is a pharmaceutical composition as defined in the second objective of the present invention, which is used to kill or inhibit the growth of persistent bacteria cells.

[0132] A sixth objective of the present invention is a kit comprising:

[0133] - a pharmaceutical composition comprising a polyamine fatty acid compound as defined in the first objective of the present invention, and

[0134] - a separate pharmaceutical composition comprising at least one antimicrobial agent.

[0135] The antimicrobial agent is as defined in the first objective of the present invention.

[0136] The present invention also relates to a method for treating bacterial diseases in a subject of need, comprising administering a pharmaceutical composition as defined in the second objective of the present invention to the subject.

[0137] The present invention also relates to a method for treating chronic infections, recurrent infections, refractory infections, persistent infections, persistent bacterial infections, persistent bacterial chronic infections, persistent bacterial recurrent infections, persistent bacterial refractory infections, or persistent bacterial infections in subjects in need, comprising administering a pharmaceutical composition as defined in the second objective of the present invention to the subject.

[0138] The present invention also relates to a method for killing or inhibiting the growth of persistent bacterial cells in subjects in need, comprising administering a pharmaceutical composition as defined in the second objective of the present invention to the subject.

[0139] The present invention also relates to the use of a pharmaceutical composition as defined in the second objective of the present invention in the preparation of a medicament for treating bacterial diseases in subjects in need.

[0140] The present invention also relates to a pharmaceutical composition as defined in the second objective of the present invention, as described herein,The invention relates to the use of pharmaceutical compositions as defined in the second objective of the invention in the preparation of medicaments for treating chronic infections, recurrent infections, refractory infections, persistent infections, persistent bacterial infections, persistent bacterial chronic infections, persistent bacterial recurrent infections, persistent bacterial refractory infections, or persistent bacterial infections in subjects in need.

[0141] The invention also relates to the use of pharmaceutical compositions as defined in the second objective of the invention in the preparation of medicaments for killing or inhibiting the growth of persistent bacterial cells in subjects in need.

[0142] The invention also relates to the use of pharmaceutical compositions as defined in the second objective of the invention for treating bacterial diseases in subjects in need.

[0143] The invention also relates to the use of pharmaceutical compositions as defined in the second objective of the invention for treating chronic infections, recurrent infections, refractory infections, persistent infections, persistent bacterial infections, persistent bacterial chronic infections, persistent bacterial recurrent infections, persistent bacterial refractory infections, or persistent bacterial infections in subjects in need, as described in CN 121443283 A (page 12 / 33 of the specification).

[0144] The present invention also relates to the use of pharmaceutical compositions as defined in the second objective of the invention for killing or inhibiting the growth of resident bacterial cells in a subject in need.

[0145] “Bacterial infection” or “bacterial disease” refers to any undesirable presence and / or growth of bacteria as pathogens in a subject. The presence of such undesirable microorganisms may have a negative impact on the health and well-being of the host subject. While the term “bacterial infection” or “bacterial disease” should not be understood to include the normal growth and / or presence of microorganisms that are normally present in a subject (e.g., in the subject’s digestive tract), it may include the pathological overgrowth of such microorganisms. “Bacterial infection” or “bacterial disease” is a pathological condition or symptom caused by the growth and / or presence of bacteria as microorganisms. Therapeutic agents for treating “bacterial infection” or “bacterial disease” are “antimicrobial” agents or “antimicrobial” drugs.

[0146] “Chronic infection,” “recurrent infection,” “refractory infection,” and “persistent infection” refer to bacterial infections that resist the host’s immune system and antibiotic treatment and are capable of reactivating as clinically significant disease with chronic symptoms.

[0147] In one embodiment, the bacterial disease is selected from cystic fibrosis, urinary tract infection, chronic otitis media, and pneumonia.

[0148] In a preferred embodiment, the bacterial disease is caused by Gram-negative bacteria.

[0149] “Object” refers to an animal, typically a warm-blooded animal, preferably a mammal. The term “mammal” herein refers to any mammal, including humans, domestic and farm animals, as well as zoo, sport, or pet animals such as dogs, cats, cattle, horses, sheep, pigs, goats, rabbits, etc. Preferably, the mammal is a primate.More preferably, it is a human being.

[0150] “Human being” means a male or female human subject at any developmental stage, including newborns, infants, toddlers, adolescents, and adults.

[0151] In one embodiment, the subject has, and preferably has been diagnosed with, a bacterial disease. In one embodiment, the subject is at risk of developing a bacterial disease. Examples of risk factors include, but are not limited to, a genetic predisposition or a family history of bacterial disease.

[0152] “Persistent bacteria” refers to a dormant variant of any type of conventional cell, particularly persistent bacteria and live unculturable cells (VBNCs). Persistent bacteria are prone to causing infectious diseases. Persistent bacteria are typically bacterial, but fungal persistent cells and yeast persistent cells are also included in this definition. Persistent bacteria represent a small subgroup of genetically identical, metabolically slow-growing cells that spontaneously enter a dormant, non-dividing state and can survive at extremely high doses of antibiotics. When the population is treated with antibiotics, conventional cells die, while persistent bacteria survive. For antibiotics to kill, an active target is required, which explains the resistance of persistent bacteria. In contrast, resistance mechanisms prevent antibiotics from binding to their targets. Drug resistance is measured by observing the ability of cells to grow in the presence of antibiotics. In most cases, the molecular mechanisms leading to persistence are unknown. As used herein, “persistent bacterial cells” refers to metabolic variants of wild-type microbial cells that are phenotypically characterized by their slow growth rate, typically 30%, 25%, 20%, 15%, 10%, 5%, or less than 5% of the growth rate of their wild-type counterparts. In some embodiments, persistent bacterial cells are dormant and, for example, show no detectable cell division within 24 hours. Furthermore, persistent bacterial cells typically form colonies that are approximately 30%, 25%, 20%, 15%, 10%, 5%, or less than 5% the size of colonies formed by their wild-type counterparts.

[0153] “Persistent bacterial-associated infection” refers to any infection involving persistent bacterial cells.

[0154] The present invention also relates to compositions comprising a polyamine fatty acid compound as defined herein and an antimicrobial agent as an anti-infective agent, preferably as an anti-persistent bacterial agent, and more preferably for non-therapeutic use in surface disinfection and / or liquid purification.

[0155] In this embodiment, the polyamined fatty acid compound is not used or applied to humans or animals. Therefore, the polyamined fatty acid compound is applied to an object, such as a medical device like a prosthesis, or to the surface of said object to disinfect it. Alternatively, the polyamined fatty acid compound is mixed with a liquid or fluid to purify it.

[0156] In this embodiment, the polyamined fatty acid compound is used in combination with an antimicrobial agent. The antimicrobial agent may be as defined in this invention.Alternatively, it may be any known preservative or reagent conventionally used for such non-therapeutic applications.

[0157] The invention also relates to polyamined fatty acid compounds corresponding to the following formula (I'), their pharmaceutically acceptable salts, and / or solvates thereof:

[0158] R1-C(=O)-NH-(CR2R3)m-[X-(CR4R5)n]p-NR6R7 (I')

[0159] wherein:

[0160] -R1 represents a straight-chain aliphatic chain containing at least 7 carbon atoms, wherein the straight-chain aliphatic chain is a monounsaturated aliphatic chain or a polyunsaturated aliphatic chain;

[0161] -R2 and R3 independently represent hydrogen atoms or C1-C8 alkyl groups in each of their m occurrences;

[0162] -R4 and R5 independently represent hydrogen atoms or C1-C8 alkyl groups in each of their n occurrences and p occurrences;

[0163] -X, in each occurrence of p, independently represents a -NR8- group or a divalent 5- to 7-membered heterocyclic alkyl group containing at least one nitrogen atom; wherein R8 represents a hydrogen atom, a C1-C6 alkyl group, or -(CH2)q-NH2, wherein q represents an integer from 1 to 5;

[0164] -R6 and R7, independently representing hydrogen atoms, C1-C8 alkyl groups, or R6 and R7 together with the nitrogen atom to which they are attached, form a 5- to 7-membered heterocyclic group optionally substituted with one to three R9s; wherein R9 represents -(CH2)u-NH2, wherein u represents an integer from 1 to 5;

[0165] -m is an integer from 2 to 10;

[0166] -n, in each occurrence of p, independently represents an integer from 1 to 5; and

[0167] -p is an integer from 0 to 4,

[0168] and excludes the following compounds and their pharmaceutically acceptable salts and / or solvates thereof:

[0169]

[0170] In formula (I'), R2, R3, R4, R5, R6, R7, m, n, and p are as defined in the first objective of the present invention. Specification 14 / 33 pages 23 CN 121443283 A

[0171] A straight-chain aliphatic chain containing at least 7 carbon atoms as the R1 group, wherein the straight-chain aliphatic chain is a monounsaturated aliphatic chain or a polyunsaturated aliphatic chain, as defined in the first objective of the present invention. More particularly, the compounds of formula (I') are selected from the following compounds (I'-a), as the seventh objective of the present invention:

[0172] Specification 15 / 33 pages 24 CN 121443283 A

[0173] Specification 16 / 33 pages 25 CN 121443283 A

[0174]

[0175] Pharmaceutically acceptable salts and / or solvates thereof.

[0176] The present invention is described in more detail in the following examples,However, this is not limited to the embodiments described. Examples

[0177] Materials

[0178] All solvents used in the synthesis were purified using standard procedures, and the reagents used in the synthesis were commercial grade. Methanol, ethyl acetate, and dichloromethane were purchased from VWR and used unpurified.

[0179] Column chromatography was performed on silica gel Macherey Nagel (70 to 230 mesh). Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker AC 300 instrument (400 MHz for 1H, 100 MHz for 13C) (using common abbreviations: s: singlet, d: doublet, t: triplet, q: quartet, m: multiplet). Tetramethylsilane was used as an internal standard. All chemical shifts are reported in ppm. Mass spectrometry analysis was performed by Spectropole (Analytical Laboratory, University of Aix-Marseille). The purity of the compounds was verified by analytical HPLC (C18 column, CH3CN:water:trifluoroacetic acid (TFA) eluent: 90:10:0.025, v / v / v) with a photodiode array (PDA) detector covering 210 nm to 310 nm, at a rate of 0.5 ml / min to 1 ml / min. All compounds showed a purity greater than 95%, as determined by HPLC-PDA at 214 nm and 254 nm. The chemical compounds synthesized in hydrochloride form were dissolved in distilled water to prepare 10 mM stock solutions.

[0180] The bacterial strains used in the following examples are the reference strains Pseudomonas aeruginosa ATCC27853, Escherichia coli ATCC25922, Klebsiella pneumoniae ATCC51296, Staphylococcus aureus

[0181] ATCC25923 and Enterococcus faecalis ATCC29212. Bacterial strains were pre-stored in 15% (v / v) glycerol at -80°C for cryoprotection. One colony of fresh culture of each strain was incubated in a culture tube containing 3 ml of Mueller-Hinton broth (MH2) and shaken overnight at 37°C. This suspension was used for inoculum preparation.

[0182] The antibiotics used in the following examples were doxycycline, erythromycin, and polymyxin B, which were purchased from Sigma. These antibiotics were dissolved in sterile distilled water or dimethyl sulfoxide (DMSO) to obtain the desired concentration.

[0183] Example 1: Preparation of various polyamine fatty acids (I) as defined in this invention

[0184] 1.1 Preparation of PAFA1

[0185] In a bicol flask, 282 mg of oleic acid (1 mmol) was placed in 15 ml of dichloromethane (CH2Cl2) with stirring.Then, 202 mg of spermine (1 mmol), 129 mg of diisopropylethylamine (1 mmol), and 442 mg of ((1H-benzo[d][1,2,3]triazol-1-yl)oxy)tris(dimethylamino)phosphonium hexafluorophosphate (((1H-Benzo[d][1,2,3]triazol-1-y1)oxy)tris(dimethylamino)phosphonium hexafluorophosphate) (1 mmol), also known as BOP, were added. The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH2Cl2 / methanol / NH4OH (7 / 3 / 1) (v / v / v). The product was obtained as a viscous yellow oil in 42% yield (cis isomer only), designated PAF1.

[0186] PAFA1 has the following chemical formula: Specification 17 / 33 Page 26 CN 121443283 A

[0187]

[0188] 1H NMR (400MHz) (MeOD) δ: 0.83 (t, J = 2.5Hz, 3H), 1.10-1.62 (m, 22H), 1.69-2.15 (m, 17H), 2.67-2.96 (m, 10H), 3.12-3.29 (m, 3H), 5.27 (m, 2H), 7.53 (m, 1H).

[0189] 13C NMR (100MHz) (MeOD) δ: 176.26, 130.88, 130.77, 50.25, 47.56, 37.94, 37.14, 33 .08, 30.86, 30.64, 30.47, 30.38, 30.34, 30.28, 29.97, 28.17, 28.01, 27.09, 23.76, 14.55.

[0190] C28H58N4O MS(ESI+)m / z=467.463(100%, [M+H]+).

[0191] 1.2 Preparation of PAFA2

[0192] In a double-necked flask, 282 mg of oleic acid (1 mmol) was placed in 15 ml of dichloromethane (CH2Cl2) with stirring, and then 131 mg of bis(3-aminopropyl)amine (1 mmol), 129 mg of diisopropylethylamine (1 mmol) and 442 mg of BOP (1 mmol) were added. The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH₂Cl₂ / methanol / NH₄OH (7 / 3 / 1) (v / v / v). The product was obtained as a viscous yellow oil in 13% yield (cis isomer only).The designation is PAFA2.

[0193] PAFA2 has the following chemical formula:

[0194]

[0195] 1H NMR (400MHz, MeOD) δ: 0.92 (t, J = 6.82Hz, 3H), 1.33 (d, J = 12.46Hz, 2OH), 1.62 (m, 2H), 1.70 (h, J = 6.99Hz, 4H), 2.06 (p, J = 5.49, 6.79Hz, 3H), 2.19 (t, J = 7.51Hz, 2H), 2.62 (m, 4H), 2.72 (t, J = 7.12Hz, 2H), 3.24 (t, J = 6.82Hz, 2H), 5.36 (t, J = 4.64Hz, 2H).

[0196] 13C NMR (100MHz) (MeOD) δ: 176.28, 130.89, 48.26, 47.85, 40.58, 38.05, 37.15, 33.21, 33.07, 30.85, 30.63, 30.46, 30.36, 30.33, 30.26, 30.23, 28.15, 27.10, 23.75, 14.50.

[0197] C24H49N3O MS (ESI+) m / z = 396.392 (100%, [M+H]+).

[0198] 1.3 Preparation of PAFA3

[0199] In a two-necked flask, 256 mg of palmitic acid (1 mmol) was added to 15 mL of dichloromethane (CH₂Cl₂) with stirring, followed by the addition of 131 mg of bis(3-aminopropyl)amine (1 mmol), 129 mg of diisopropylethylamine (1 mmol), and 442 mg of BOP (1 mmol). The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH₂Cl₂ / methanol / NH₄OH (7 / 3 / 1) (v / v / v). The product was given as a viscous yellow oil in 13% yield and designated PAFA₃.

[0200] PAFA3 has the following chemical formula: Specification 18 / 33 page 27 CN 121443283 A

[0201]

[0202] 1H NMR (400MHz, MeOD) δ: 0.92 (t, J = 6.81Hz, 3H), 1.31 (s, 24H), 1.62 (dt, J = 8.31, 14.76Hz, 2H), 1.71 (dt, J = 6.76, 14.14Hz, 4H), 2.19 (t, J = 7.49Hz, 2H), 2.62 (m, 4H), 2.72 (t, J = 7.11Hz, 2H), 3.24 (t, J = 6.81Hz,2H).

[0203] 13C NMR (100MHz, MeOD) δ: 176.34, 48.26, 47.85, 40.58, 38.05, 37.16, 33.15, 33.09, 30.81, 30.78, 30.75, 30.67, 30.49, 30.47, 30.34, 30.23, 27.11, 23.75, 14.47.

[0204] C22H47N3O MS (ESI+) m / z = 370.375 (100%, [M+H]+).

[0205] 1.4 Preparation of PAFA4

[0206] In a two-necked flask, 256 mg of palmitic acid (1 mmol) was added to 15 mL of dichloromethane (CH₂Cl₂) with stirring, followed by the addition of 202 mg of spermine (1 mmol), 129 mg of diisopropylethylamine (1 mmol), and 442 mg of BOP (1 mmol). The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH₂Cl₂ / methanol / NH₄OH (7 / 3 / 1) (v / v / v). The product was obtained as a viscous yellow oil in 51% yield and designated PAFA₄.

[0207] PAFA4 has the following chemical formula:

[0208]

[0209] 1H NMR (400MHz, MeOD) δ: 0.87 (m, 4H), 1.25 (s, 32H), 1.62 (s, 4H), 1.72 (p, J = 6.40Hz, 3H), 2.15 (m, 4H), 2.67 (s, 3H), 2.75 (t, J = 6.55Hz, 3H), 3.33 (q, J = 5.93Hz, 3H).

[0210] 13C NMR (100MHz, MeOD) δ: 175.69, 49.74, 47.01, 37.44, 36.88, 32.43, 30.18, 30.16, 30.05, 29.88, 29.86, 29.82, 29.33, 27.59, 26.50, 23.15, 14.30.

[0211] C26H56N4O MS(ESI+) m / z=441.446(100%, [M+H]+).

[0212] 1.5 Preparation of PAFA5

[0213] In a double-necked flask, 328 mg of docosahexanoic acid (1 mmol) was placed in 15 ml of dichloromethane (CH2Cl2) with stirring.Then, 131 mg of bis(3-aminopropyl)amine (1 mmol), 129 mg of diisopropylethylamine (1 mmol), and 442 mg of BOP (1 mmol) were added. The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH2Cl2 / methanol / NH4OH (7 / 3 / 1) (v / v / v). The product was given as a viscous yellow oil in 68% yield (only one isomer with all double bonds in the cis configuration), designated PAFA5.

[0214] PAFA5 has the following chemical formula:

[0215]

[0216] 1H NMR (400MHz, MeOD) δ: 0.99 (t, J = 7.54Hz, 3H), 1.72 (m, 5H), 2.10 (p, J = 7.33Hz, 3H), 2.24 (t, J = 7.38Hz, 2H), 2.40 (q, J = 6.89Hz, 2H), 2.62 (dt, J = 7.27, 10.40Hz, 4H), 2.73 (t, J = 7.10Hz, 2H), 2.87 (m, 11H), 3.23 (d, J = 6.81Hz, 2H), 5.38 (m, 13H).

[0217] 13C NMR (100MHz, MeOD) δ: 175.42, 132.81, 130.18, 129.47, 129.25, 129.20, 129.15, 129.10, 128.92, 128.18, 48.23, 47.82, 40.53, 38.10, 36.95, 32.91, 30.18, 26.57, 26.52, 26.44, 24.74, 21.51, 14.71.

[0218] C28H47N3O MS (ESI+) m / z = 442.386 (100%, [M+H]+).

[0219] 1.6 Preparation of PAFA6

[0220] In a double-necked flask, 328 mg of docosahexanoic acid (1 mmol) was placed in 15 mL of dichloromethane (CH2Cl2) with stirring, followed by the addition of 202 mg of spermine (1 mmol), 129 mg of diisopropylethylamine (1 mmol), and 442 mg of BOP (1 mmol). The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH2Cl2 / methanol / NH4OH (7 / 3 / 1) (v / v / v). The product was obtained as a viscous yellow oil.The yield was 39% (only one isomer with all double bonds in the cis configuration), and it was designated PAFA6.

[0221] PAFA6 has the following chemical formula:

[0222]

[0223] 1H NMR (400MHz, MeOD) δ: 0.72 (t, J = 2.5Hz, 3H), 1.07-1.53 ​​(m, 10H), 1.84-1.98 (m, 3H), 1.98-2.18 (m, 3H), 2.37-2.46 (m, 8H), 2.52-2.70 (m, 16H), 3.03 (m, 3H), 3.45 (s, 2H), 4.82 (s, 1H), 5.21-5.06 (m, 12H), 8.02 (m, 1H).

[0224] 13C NMR (100MHz, MeOD) δ: 175.36, 132 .79, 130 .15, 129 .46, 129 .25, 129 .18, 129.13, 129.09, 128.90, 128.16, 50.26, 47.58, 38.01, 36.93, 29.98, 28.02, 26.57, 26.52, 26.45, 24.72, 21.51, 14.76.

[0225] C32H56N4O MS(ESI+) m / z=513.455(100%, [M+H]+).

[0226] 1.7 Preparation of PAFA7

[0227] In a two-necked flask, 280 mg of linoleic acid (1 mmol) was added to 15 mL of dichloromethane (CH₂Cl₂) with stirring, followed by the addition of 74 mg of 1,3-propanediamine (1 mmol), 129 mg of diisopropylethylamine (1 mmol), and 442 mg of BOP (1 mmol). The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH₂Cl₂ / methanol / NH₄OH (7 / 3 / 1) (v / v / v). The product was obtained as a viscous yellow oil in 71% yield, as a mixture of isomers, designated PAFA7.

[0228] PAFA7 has the following chemical formula:

[0229]

[0230] 1H NMR (400MHz, MeOD) δ: 0.94 (t, J = 6.90Hz, 4H), 1.36 (s, 17H), 1.63 (m, 3H), 1.69 (m, 2H), 2.09 (q, J = 6.59Hz, 4H), 2.21 (m, 2H), 2.70 (t, J = 6.98Hz, 2H), 2.80 (t, J = 6.30Hz, 2H), 3.26 (t, J = 6.79Hz, 2H), 5.37 (d, J = 44.91Hz,4H). Specification 20 / 33 pages 29 CN 121443283 A

[0231] 13C NMR (100MHz, MeOD)δ: 176.31, 130.93, 130.86, 129.10, 129.04, 39.52, 37.45, 37.12, 32.96, 32.66, 30.74, 30.47, 30.37, 30.32, 30.27, 28.18, 27.07, 26.55, 23.63, 14.51.

[0232] C21H40N2O MS (ESI+) m / z=337.562 (100%, [M+H]+).

[0233] 1.8 Preparation of PAFA8

[0234] In a double-necked flask, 280 mg of linoleic acid (1 mmol) was placed in 15 mL of dichloromethane (CH2Cl2) with stirring, followed by the addition of 88 mg of putrescine (1 mmol), 129 mg of diisopropylethylamine (1 mmol), and 442 mg of BOP (1 mmol). The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH2Cl2 / methanol fNH4OH (7 / 3 / 1) (v / v / v). The product was obtained as a viscous yellow oil in 63% yield (only one isomer with all double bonds in the cis configuration), designated PAFA8.

[0235] PAFA8 has the following chemical formula:

[0236]

[0237] 1H NMR (400MHz, MeOD) δ: 0.91 (t, J = 6.87Hz, 3H), 1.34 (s, 15H), 1.54 (p, J = 3.48Hz, 4H), 1.61 (m, 3H), 2.07 (q, J = 6.61Hz, 4H), 2.18 (t, J = 7.53Hz, 2H), 2.73 (t, J = 6.63Hz, 2H), 2.78 (t, J = 6.23Hz, 2H), 3.18 (t, J = 6.37Hz, 2H), 5.35 (d, J = 44.70Hz, 4H).

[0238] 13C NMR (100MHz, MeOD) δ: 176.28, 130.95, 130.87, 129.11, 129.04, 41.64, 39.94, 37.16, 32.66, 30.73, 30.47, 30.34, 30.32, 30.25, 29 .65, 28.16, 27.72, 27.08, 26.54, 23.62, 14.45.

[0239] C22H42N2O MS (ESI+) m / z=351.328 (100%,[M+H]+) .

[0240] 1.9 Preparation of PAFA9

[0241] In a double-necked flask, 280 mg of linoleic acid (1 mmol) was placed in 15 ml of dichloromethane (CH2Cl2) with stirring, followed by the addition of 145 mg of 3,3-diamino(N-methyldipropylamine) (1 mmol), 129 mg of diisopropylethylamine (1 mmol), and 442 mg of BOP (1 mmol). The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH2Cl2 / methanol / NH4OH (7 / 3 / 1) (v / v / v). The product was obtained as a viscous yellow oil in 48% yield (only one isomer with all double bonds in the cis configuration), designated PAFA9.

[0242] PAFA9 has the following chemical formula:

[0243]

[0244] 1H NMR (400MHz, MeOD) δ: 0.87 (t, J = 6.85Hz, 3H), 1.29 (s, 15H), 1.57 (m, 2H), 1.76 (p, J = 6.84Hz, 2H), 1.89 (p, J = 7.09Hz, 2H), 2.02 (q, J = 6.62Hz, 4H), 2.17 (m, 2H), 2.46 (s, 3H), 2.68 (m, 2H), 2.73 (t, J = 6.34Hz, 2H), 2.78 (t, J = 7.18Hz, 2H), 3.02 (t, J = 7.18Hz, 2H). .07Hz, 2H), 3.20 (t, J=6.74Hz, 2H), 5.30 (d, J=44.49Hz, 4H).

[0245] 13C NMR (100MHz, MeOD) δ: 176.69, 130.85, 130.80, 128.97, 128.94, 55.67, 41.09, 39.52, 37.64, 36.99, 32.53, 30.63, 30.35, 30.23, 30 .22, 30.17, 28.08, 28.06, 26.89, 26.78, 26.45, 24.18, 23.51, 14.41.

[0246] C25H49N3O MS(ESI+) m / z=408.390(100%, [M+H]+).

[0247] 1.10 Preparation of PAFA10

[0248] In a double-necked flask, 280 mg of linoleic acid (1 mmol) was placed in 15 ml of dichloromethane (CH2Cl2) with stirring.Then, 188 mg of tris(3-aminopropyl)amine (1 mmol), 129 mg of diisopropylethylamine (1 mmol), and 442 mg of BOP (1 mmol) were added. The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH2Cl2 / methanol / NH4OH (7 / 3 / 1) (v / v / v). The product was given as a viscous yellow oil in 28% yield (only one isomer with all double bonds in the cis configuration), designated PAFA10.

[0249] PAFA10 has the following chemical formula:

[0250]

[0251] 1H NMR (400MHz, MeOD) δ: 0.89 (t, J = 6.90Hz, 3H), 1.32 (s, 16H), 1.64 (d, J = 56.06Hz, 10H), 2.05 (q, J = 6.60Hz, 4H), 2.16 (t, J = 7.52Hz, 2H), 2.48 (dt, J = 7.34, 14.36Hz, 6H), 2.75 (q, J = 6.85Hz, 6H), 3.17 (t, J = 7.00Hz, 2H), 5.33 (d, J = 44.88Hz, 4H).

[0252] 13C NMR (100MHz, MeOD) δ: 176.19, 130.95, 130.86, 129.12, 129.04, 52.78, 52.48, 40.70, 38.65, 37.18, 32.66, 30.74, 30.47, 30.37, 30.33, 30.27, 29.27, 28.16, 27.65, 27.10, 26.54, 23.62, 14.47.

[0253] C27H54N4O MS (ESI+) m / z = 451.442 (100%, [M+H]+).

[0254] 1.11 Preparation of PAFA11

[0255] In a two-necked flask, 280 mg of linoleic acid (1 mmol) was added to 15 mL of dichloromethane (CH₂Cl₂) with stirring, followed by the addition of 131 mg of bis(3-aminopropyl)amine (1 mmol), 129 mg of diisopropylethylamine (1 mmol), and 442 mg of BOP (1 mmol). The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH₂Cl₂ / methanol / NH₄OH (7 / 3 / 1) (v / v / v). The product was given as a viscous yellow oil in 52% yield (only one isomer with all double bonds in the cis configuration).The part numbered PAFA11.

[0256] PAFA11 has the following chemical formula: Specification 22 / 33 pages 31 CN 121443283 A

[0257]

[0258] 1H NMR (400MHz, MeOD) δ: 0.91 (m, 3H), 1.35 (m, 15H), 1.62 (q, J = 7.43Hz, 3H), 1.73 (dp, J = 7.06, 13.90Hz, 4H), 2.07 (dt, J = 6.19, 8.38Hz, 4H), 2.19 (t, J = 7.50Hz, 2H), 2.64 (t, J=7.13Hz, 2H), 2.70 (t, J=7.11Hz, 2H), 2.78 (t, J=6.17Hz, 2H), 2.84 (t, J=7.07Hz, 2H), 3.23 (t, J=6.79Hz, 2H), 5.34 (m, 4H).

[0259] 13C NMR (100MHz, MeOD) δ: 176.45, 130.95, 130.86, 129.12, 129.04, 48.15, 47.56, 40.29, 37.85, 37.13, 32.66, 30.76, 30.73, 30.47, 30 .35, 30 .32, 30 .26, 30 .01, 28 .16, 27.09, 26.53, 23.62, 14.45.

[0260] C24H47N3O MS(ESI+) m / z=394.378(100%, [M+H]+).

[0261] 1.12 Preparation of PAFA12

[0262] In a double-necked flask, 280 mg of linoleic acid (1 mmol) was placed in 15 ml of dichloromethane (CH2Cl2) with stirring, and then 146 mg of triethylenetetramine (1 mmol), 129 mg of diisopropylethylamine (1 mmol) and 442 mg of BOP (1 mmol) were added. The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH2Cl2 / methanol / NH4OH (7 / 3 / 1) (v / v / v). The product was obtained as a viscous yellow oil in 47% yield (only one isomer with all double bonds in the cis configuration), designated PAFA12.

[0263] PAFA12 has the following chemical formula:

[0264]

[0265] 1H NMR (400MHz, MeOD) δ: 0.93 (t, J = 6.87Hz, 3H), 1.35 (s, 16H), 1.62 (m, 2H), 2.08 (q, J = 6.60Hz, 4H), 2.21 (t, J = 7.54Hz, 2H), 2.74 (d,J=13.16Hz, 12H), 3.33 (d, J=12.75Hz, 4H), 5.36 (d, J=45.03Hz, 4H).

[0266] 13C NMR (100MHz, MeOD) δ: 176.61, 130.95, 130.87, 129.11, 129.04, 51.69, 49.34, 41.47, 39.86, 37.15, 32.64, 30.71, 30.45, 30.33, 30 .31, 30.24, 28.15, 26.97, 26.52, 23.61, 14.44.

[0267] C24H48N4O MS(ESI+) m / z = 409.386 (100%, [M+H]+).

[0268] 1.13 Preparation of PAFA13

[0269] In a double-necked flask, 280 mg of linoleic acid (1 mmol) was placed in 15 ml of dichloromethane (CH2Cl2) with stirring, and then 188 mg of N,N'-bis(3-aminopropyl)-1,3-propanediamine (1 mmol), 129 mg of diisopropylethylamine (1 mmol) and 442 mg of BOP (1 mmol) were added. The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH2Cl2 / methanol / NH4OH (7 / 3 / 1) (v / v / v). The product was obtained as a viscous yellow oil in 35% yield (only one isomer with all double bonds in cis configuration), designated PAFA13.

[0270] PAFA13 has the following chemical formula:

[0271]

[0272] 13C NMR (100MHz, D2O) δ: 176.78, 130.79, 130.64, 128.83, 128.69, 45.67, 45.55, 37.52, 36.93, 32.35, 30.66, 30.22, 28.14, 27.97, 26.80, 26.69, 26.44, 24.70, 23.83, 23.62, 23.42, 14.77.

[0273] C27H54N4O MS (ESI+) m / z = 451.453 (100%, [M+H]+).

[0274] 1.14 Preparation of PAFA14

[0275] In a double-necked flask, 280 mg of linoleic acid (1 mmol) was placed in 15 ml of dichloromethane (CH2Cl2) with stirring.Then, 202 mg spermine (1 mmol), 129 mg diisopropylethylamine (1 mmol), and 442 mg BOP (1 mmol) were added. The mixture was stirred at room temperature for 12 hours and then concentrated under vacuum. The crude mixture was purified by silica gel chromatography using methanol as eluent, followed by a mixture of CH2Cl2 / methanol / NH4OH (7 / 3 / 1) (v / v / v). The product was given as a viscous yellow oil in 33% yield (only one isomer with all double bonds in the cis configuration), designated PAFAv4.

[0276] PAFA14 has the following chemical formula:

[0277]

[0278] 1H NMR (400MHz, MeOD) δ: 0.92 (t, J = 8.0Hz, 3H), 1.27–1.35 (m, 14H), 1.54–1.73 (m, 13H), 2.05–2.09 (m, 4H), 2.17–2.21 (t, J = 8.0Hz, 2H), 2.31–2.36 (m, 3H), 2.60–2.63 (m, 6H), 2.78–2.80 (t, J = 8.0Hz, 4H), 3.22–3.25 (m, 2H), 3.32–3.40 (m, 3H), 5.30–5.4 (m, 3H)

[0279] 13C NMR (100MHz, MeOD) δ: 176.36, 130.96, 130.87, 129.13, 129.05, 69.79, 56.12, 56.08, 53.67, 50.39, 47.71, 45.52, 37.99, 37.14, 32

[0280] C28H56N4O MS(ESI+)m / z=465.465(100%, [M+H]+).

[0281] Example 2: Intrinsic antibacterial activity of polyamined fatty acids (I) and their toxicity to eukaryotic cells

[0282] The polyamined fatty acid compounds obtained by the reductive amination reaction described in Example 1 above were prepared in the form of hydrochloride for biological testing.

[0283] The antibacterial activity of the polyamined fatty acid compounds was measured using the standard microdilution method based on the Clinical and Laboratory Standards Institute (CLSI) guidelines. This method was slightly modified by increasing the assay volume to 200 μl to improve reproducibility. Specification 24 / 33 pages 33 CN 121443283 A

[0284] 2.1 Preparation of preculture

[0285] A negative control was prepared,This corresponds to 2 ml of sterile culture medium, and the positive control corresponds to a mixture of 1980 μl of culture medium and 20 μl of bacterial suspension from thawed biological strains (the biological strains were stored in glycerol at -80°C).

[0286] The test tubes were incubated at 100 rpm in an Infors shaking incubator at 37°C for 24 hours.

[0287] The bacteria were handled under a laboratory fume hood, with a UV cycling procedure performed before any handling, and only sterile materials were used.

[0288] The polyamine fatty acid compounds to be tested were prepared in water at a concentration of 10 mM.

[0289] 2.2 Preparation of 96-well microplates and measurement of intrinsic antimicrobial activity

[0290] The required volume of the microbial suspension to be inoculated was calculated such that the optical density (OD) of each well, measured at 600 nm using a spectrophotometer, corresponds to a value equal to 0.01. In the microplate, the first row corresponds to the negative control (200 μl of sterile medium per well), the second row corresponds to the positive control (inoculated medium), and the third row contains bacterial suspension and 8 μl of the given polyamine fatty acid compound added twice to each well. Subsequently, a half-fold serial dilution is performed starting from this row. The first column serves as an inhibition control. A sterile filter membrane is then placed on the microplate, allowing gas to pass through but not contaminants. The microplate is incubated at 37°C in a humid atmosphere for 24 hours. After 24 hours of incubation, optical density is measured by diluting 100 μl of bacterial suspension in 900 μl of sterile medium.

[0291] The medium used is Mueller-Hinton medium (MH) for bacteria. All tests are performed twice.

[0292] 2.3 Toxicity of Polyamine Fatty Acid Compound (I) to Mammalian Cells

[0293] The cytotoxic activity of the polyamine fatty acid compound was measured using the WST1 assay. This is a colorimetric assay for measuring viability and cell proliferation rate. It is based on the cleavage of colorless tetrazolium salt WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetraazolium]-1,3-benzenedisulfonate) into a yellow formazan derivative by mitochondrial dehydrogenase, which can be quantified by spectrophotometry at 420 nm to 480 nm.

[0294] WST1 assay was performed on Chinese hamster ovary cells. CHO-K1 cells (ATCC, USA) were maintained in Mac Coy's 5A medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine and streptomycin penicillin mixture (100 U / ml: 10 μg / ml). Incubation was performed at 37°C in a CO2-rich atmosphere (5% v / v) and passaged every two days.

[0295] Cells were transferred to 96-well plates (25,000 cells / ml) and placed in complete Mac Coy's 5A medium.The cells were maintained at 37°C in a humid, CO2-rich atmosphere (5% vol%) for 24 hours. In two assays, a given polyamined fatty acid compound at an increased concentration was added to each well, with each assay run including eight growth controls containing only cells in culture medium. After 24 hours at 37°C (5% CO2), the culture medium was removed, the cells were washed in phosphate-buffered saline (PBS), and 50 μl of PBS containing 10% WST1 reagent was added to each well. After incubation at 37°C for 20 minutes, the results were read spectrophotometrically at 450 nm.

[0296] The results are presented as dose-response relationships and modeled using TableCurve software via nonlinear regression analysis. The 50% inhibitory concentration (IC50) represents the concentration of the polyamined fatty acid compound that reduces cell viability to 50%.

[0297] Table 1 below reports the intrinsic antibacterial activity (MIC, in μM) of some of the polyamined fatty acids prepared in Example 1 and their toxicity to eukaryotic cells (IC50, in μM). Instructions for Use, Pages 25 / 33, 34, CN 121443283 A

[0298]

[0299] ND: Not determined

[0300] Table 1

[0301] Table 1 shows that the polyamined fatty acid compound as defined in this invention does not exhibit intrinsic antibacterial activity against one or more of the following bacterial strains: Pseudomonas aeruginosa ATCC27853, Enterococcus faecalis ATCC29212, Staphylococcus aureus ATCC25923, Escherichia coli ATCC25922.

[0302] Furthermore, this polyamined fatty acid compound (I) has been shown to be non-toxic to mammalian cells.

[0303] Example 3: Use of polyamined fatty acid compound (I) as defined in this invention in combination with an antibacterial agent

[0304] 100 μl of liquid culture medium was placed in each well of a 96-well microplate and then inoculated with the bacterial suspension prepared as a positive control in Example 2.1. The required inoculation volume was calculated to achieve an OD of 0.01, corresponding to approximately 5 × 10⁶ bacteria per well. In the plate, the first row corresponds to the negative control (200 μl of sterile medium per well), the second row to the positive control (100 μl of sterile medium plus 100 μl of bacterial suspension), and the third row contains 192 μl of medium and 8 μl of the polyamine fatty acid compound to be tested per well. Serial dilutions were then performed starting from this row. Next, 3 μl to 8 μl of doxycycline solution (1 mg dissolved in 20 ml) was added to each well in this row to obtain a final antibiotic concentration of 2 μg / ml. Then, 92 μl of bacterial suspension was added to rows 3 through 8. After incubation at 37°C under a humid atmosphere for 24 hours…The MIC was determined in the presence of X μg / ml polyamined fatty acid compounds and an added concentration of 2 μg / ml doxycycline. After incubation at 37°C for 24 hours, 40 μl of nitrotetrazolium iodide was added to each well to indicate the presence of viable bacteria by staining the medium pink.

[0305] Table 2 below reports the antibacterial activity (MIC, in μM) of some polyamined fatty acids prepared in Example 1 in combination with the two antibacterial agents doxycycline and erythromycin. For the Gram-negative strain Pseudomonas aeruginosa (ATCC27853), doxycycline had an MIC of 32 μg / ml. For the Gram-negative strain Escherichia coli (ATCC28922), erythromycin had an MIC of 50 μg / ml. Instructions for Use, Pages 26 / 33, 35, CN 121443283 A

[0306]

[0307] ND: Not determined

[0308] Table 2

[0309] In fact, Table 2 shows the ability of the polyamined fatty acid (I) prepared in Example (I) to enhance the ability of doxycycline and / or erythromycin used at a concentration of 2 μg / ml against Pseudomonas aeruginosa bacterial strain ATCCC27853 and Escherichia coli bacterial strain ATCC25922, which are known Gram-negative resistant strains.

[0310] Example 4: Use of polyamined fatty acid compound (I) as a enhancer of known antibiotics as defined in this invention

[0311] 100 μl of liquid culture medium was added to each well of a 96-well microplate and then inoculated with the bacterial suspension prepared as a positive control in Example 2.1. The volume to be inoculated was calculated to make the OD 0.01, which corresponds to approximately 5 × 10⁶ bacteria per well. In this plate, the first row corresponds to the negative control (200 μl of sterile culture medium per well), the second row corresponds to the positive control (100 μl of sterile culture medium plus 100 μl of bacterial suspension per well), and the third row contains 192 μl of culture medium and 8 μl of the antibiotic solution to be tested (10 mg / ml) per well. Serial dilutions are then performed starting from this row. Next, 10 μl of fatty acid solution (final concentration 10 μM) is added to each well in this row. Then, 90 μl of bacterial suspension is added to rows 3 through 8. After incubation at 37°C under a humid atmosphere for 24 hours, the MIC of the test antibiotic is determined. After incubation at 37°C for 24 hours, 40 μl of nitrotetrazoleonium iodide is added to each well.The presence of live bacteria was indicated by staining the culture medium pink.

[0312] Table 3 below reports the antimicrobial activity (MIC, in μg / mL) of some polyamine fatty acids prepared in Example 1 with various antimicrobial agents: doxycycline (DOX), tetracycline (TET), oxytetracycline (OXY) and minocycline (MINO) against various Pseudomonas aeruginosa bacterial strains (PA01, PA0509 and PA263) and Escherichia coli bacterial strains (AG-100 and AG-100A). Specification 27 / 33 pages 36 CN 121443283 A

[0313]

[0314] Table 3

[0315] Table 3 shows that, by combining polyamine fatty acid compound (I) with antimicrobial agents, the MIC against Gram-negative bacterial strains is significantly reduced (at least by 1 / 4).

[0316] This demonstrates the enhancing effect of polyamine fatty acid compound (I) as defined in this invention.

[0317] Example 5: Preparation of other polyamined fatty acid compounds (I) as defined in this invention and their use as enhancers of known antibiotics

[0318] Several other polyamined fatty acid compounds PAFA15 to PAFA27 corresponding to formula (I) have been prepared according to the same procedure as described for compounds PAFA1 to PAFA14.

[0319] The structures of polyamined fatty acid compounds PAFA15 to PAFA27 and their corresponding NMR and mass spectrometric characteristics are reported below.

[0320] PAFA15 has the following chemical formula:

[0321]

[0322] 1H NMR (400MHz, MeOD) δ 5.36 (m, 13H), 4.87 (s, 4H), 3.23 (t, J = 6.77Hz, 2H), 2.85 (dt, J = 5.96, 11.87Hz, 11H), 2.81 (d, J = 4.81Hz, 2H), 2.72 (d, J = 17.52Hz, 4H), 2.64 (m, 2H), 2.38 (m, 2H), 2.23 (t, J = 7.22Hz, 2H), 2.07 (qd, J = 1.34, 7.41Hz, 2H), 1.71 (m, 4H), 0.97 (t, J=7.55Hz, 3H).

[0323] 13C NMR (101MHz, MeOD) δ 175.52, 132.81, 130.19, 129.47, 129.25, 129.21, 129.18, 129.14, 129.10, 128.92, 128.18, 49.29, 48.17, 47.63, 40.30, 37.98, 36.95, 31.15, 30.10, 26.58, 26.56, 26.53, 26.44, 24.74, 21.51,14.69. Specification 28 / 33 Page 37 CN 121443283 A

[0324] C30H52N4O MS(ESI+) m / z=485.4172(100%, [M+H]+).

[0325] PAFA16 has the following chemical formula:

[0326]

[0327] 1H NMR (400MHz, MeOD)δ: 3.24 (t, J=6.79Hz, 2H), 2.82 (q, J=7.40Hz, 2H), 2.67 (dt, J=7.14, 23.60Hz, 4H), 2.19 (t, J=7.47Hz, 2H), 1.72 (dq, J=6.99, 11.81Hz, 4H), 1.61 (t, J=7.29Hz, 2H), 1.31 (d, J=10.00Hz, 18H), 0.91 (t, J=6.76Hz, 3H).

[0328] 13C NMR (101MHz, MeOD) δ: 176.50, 69.85, 53.78, 48.16, 47.58, 45 .51, 40.31, 38.65, 37.87, 37.15, 33.06, 30.97, 30.73, 30.64, 30.46, 30.33, 30.30, 30.02, 27.09, 26.88, 23.73, 14.45.

[0329] C18H39N3O MS(ESI+) m / z=314.3127(100%, [M+H]+).

[0330] PAFA17 has the following chemical formula:

[0331]

[0332] 1H NMR (400MHz, MeOD) δ: 3.25(t, J=6.78Hz, 3H), 2.78(dt, J=7.11, 23.17Hz, 2H), 2.67(m, 6H), 2.20(t, J=7.49Hz, 3H), 1.75(h, J=6.78Hz, 4H), 1.62(dq, J=3.55, 7.03Hz, 7H), 1.33 (d, J=10.55Hz, 21H), 0.92 (m, 4H).

[0333] 13C NMR (101MHz, MeOD) δ: 176.43, 50.12, 50.06, 47.98, 47.47, 40.35, 37.89, 37.14, 33.07, 31.37, 30.75, 30.66, 30.47, 30.34, 29.89, 29.84, 28.09, 27.87, 27.78, 27.10, 25.32, 23.74,14.49.

[0334] C22H48N4O MS(ESI+) m / z=385.3845(100%, [M+H]+).

[0335] PAFA18 has the following chemical formula:

[0336]

[0337] 1H NMR (400MHz, MeOD)δ: 5.34(dh, J=4.95, 11.28Hz, 6H), 3.45(d, J=6.48Hz, 2H), 3.25(t, J=6.80Hz, 3H), 2.80(m, 13H), 2.12(dq, J=7.20, 46.41Hz, 7H), 1.67(m, 12H), 1.32(s, 10H), 0.97(m, 2H). 13C NMR (101MHz, MeOD) δ: 176.73, 132.72, 131.05, 129.20, 129.18, 128.85, 128.22, 69.54, 53.41, 49.48, 46.88, 45.35, 37

[0339] C28H54N4O MS(ESI+) m / z = 463.4329 (100%, [M+H]+).

[0340] PAFA19 has the following chemical formula: Specification 29 / 33 pages 38 CN 121443283 A

[0341]

[0342] 1H NMR (400MHz, MeOD) δ: 5.36 (m, 7H), 3.40 (d, J = 28.64Hz, 2H), 3.25 (t, J = 6.76Hz, 2H), 2.78 (m, 12H), 2.16 (m, 8H), 1.77 (ddt, J = 7.01, 9.25, 14.07Hz, 3H), 1.63 (m, 7H), 1.36 (m, 9H), 0.91 (t, J=6.73Hz, 3H).

[0343] 13C NMR (101MHz, MeOD) δ: 176.43, 131.15, 130.63, 129.26, 129.17, 129.05, 128.73, 74 .26, 69.53, 55.77, 53.47, 49.85, 47.67, 47.22, 45.31, 40.05, 37.74, 36.98, 32.58, 30.38, 30.17, 29.42, 28.13, 27.93, 27.49, 26.68,25.03, 23.57, 14.50.

[0344] C28H54N4O MS(ESI+) m / z = 463.4246 (100%, [M+H]+).

[0345] PAFA20 has the following chemical formula:

[0346]

[0347] 1H NMR (400MHz, MeOD) δ: 3.23 (t, J = 6.80Hz, 4H), 2.65 (m, 8H), 2.19 (t, J = 7.47Hz, 3H), 1.63 (m, 11H), 1.32 (m, 20H), 0.91 (m, 4H).

[0348] 13C NMR (101MHz, MeOD) δ: 176.41, 50.30, 50.26, 48 .09, 47.61, 40.47, 37.97, 37.95, 37.15, 33.06, 32.14, 30.70, 30.66, 30.46, 30.33, 30.05, 30.03, 28.07, 28.05, 27.96, 27.11, 23.73, 14.47.

[0349] C21H46N4O MS(ESI+) m / z=371.3754(100%, [M+H]+).

[0350] PAFA21 has the following chemical formula:

[0351]

[0352] 13C NMR (101MHz, MeOD)δ:176.41, 48.21, 47

[0353] 1H NMR (400MHz, MeOD) δ: 3.24 (t, J=6.82Hz, 2H), 2.76 (t, J=7.11Hz, 2H), 2.64 (m, 4H), 2.19 (t, J=7.50Hz, 2H), 1.71 (p, J=7.32Hz, 4H), 1.61 (m, 3H), 1.32 (m, 16H), 0.90 (m, 3H).

[0354] C17H37N3O MS(ESI+) m / z=300.2934(100%, [M+H]+).

[0355] PAFA22 has the following chemical formula:

[0356] Specification 30 / 33 pages 39 CN 121443283 A

[0357] 1H NMR (400MHz, MeOD) δ: 5.38(m, 5H), 3.38(s, 5H), 3.24(dt, J=6.86, 12.02Hz, 2H), 2.83(m, 5H), 2.67(dt, 2H), 2.83(m, 5H), 2.67(dt,J=7.20, 18.26Hz, 3H), 2.35 (m, 1H), 2.16 (m, 6H), 1.70 (ddq, J= 7.37, 15.δ9, 34.18Hz, 6H), 1.36 (m, 8H), 0.93 (t, J=6.73Hz, 3H).

[0358] 13C NMR (101MHz, MeOD) δ: 176.37, 176.18, 131.16, 130.64, 130.62, 129.27, 129.19, 129.05, 128.74, 74.26, 69.71, 53.72, 53.59, 49.85, 48.08, 47.58, 45.39, 40.29, 38.66, 37.93, 37.03, 37.01, 32.59, 31 .44, 30.38, 30.28, 29.96, 28.12, 27.92, 27.32, 26.79, 26.72, 26.51, 23.57, 14.45.

[0359] C24H45N3O MS (ESI+) m / z=392.3596 (100%, [M+H]+).

[0360] PAFA23 has the following chemical formula:

[0361]

[0362] 1H NMR (400MHz, MeOD) δ: 5.33 (hept, J = 6.40, 7.12Hz, 14H), 3.24 (q, J = 3.35, 4.86Hz, 3H), 2.80 (dq, J = 5.41, 5.87, 26.52Hz, 16H), 2.58 (m, 7H), 2.36 (m, 3H), 2.22 (m, 3H), 2.06 (p, J = 7.45Hz, 3H), 1.42 (d, J = 4.14Hz, 1H), 0.95 (t, J = 7.54Hz, 4H).

[0363] 13C NMR (101MHz, MeOD) δ: 175 .56, 132 .80, 130 .14, 129 .45, 129 .27, 129 .20, 129.18, 129.13, 129.08, 128.90, 128.16, 56.07, 55.17, 39.63, 38.53, 36.93, 28.79, 26.59, 26.56, 26.55, 26.51, 26.45, 26.43, 24.63, 21.50, 14.74, 14.70.

[0364] C31H54N4O MS(ESI+)m / z=499.4331(100%, [M+H]+).

[0365] PAFA24 has the following chemical formula:

[0366]

[0367] 1H NMR (400MHz,MeOD) δ: 5.34 (ttd, J=1.41, 5.70, 6.16, 11.65Hz, 6H), 3.18 (t, J =7.03Hz, 2H), 2.81 (dt, J=6.50, 10.02Hz, 8H), 2.49 (dt, J=7.41, 22.06Hz, 6H), 2.10 (m, 7H), 1.66 (m, 9H), 1.33 (m, 11H), 0.96 (t, J=7.53Hz, 3H) .

[0368] 13C NMR (101MHz, MeOD) δ: 176.25, 132.73, 131.06, 130.94, 130.85, 129.20, 129 .11, 129.03, 128.86, 128.23, 125.14, 124.98, 52.67, 52.34, 40.44, 38.55, 37.18, 32.65, 30.71, 30.47, 30.36, 30.33, 30.26, 28.17, 28.08, 27.70, 27.09, 26.85, 26.54, Specification 31 / 33 pages 40 CN 121443283 A 26.52, 26.40, 23.62, 21.49, 14.68, 14.45.

[0369] C27H52N4O MS (ESI+) m / z = 449.4175 (100%, [M+H]+).

[0370] PAFA25 has the following chemical formula:

[0371]

[0372] 1H NMR (400MHz, MeOD) δ: 5.35 (m, 6H), 3.25 (m, 4H), 2.77 (ddt, J = 7.17, 31.57, 64.30Hz, 9H), 2.12 (m, 7H), 1.71 (m, 7H), 1.35 (p, J = 4.87Hz, 10H), 0.98 (t, J = 7.55Hz, 3H).

[0373] 13C NMR (101MHz, MeOD) δ: 176.53, 132.73, 131 .05, 130.94, 130.85, 129.20, 129.03, 128.87, 128.23, 48.09, 47.43, 40.14, 37.77, 37.13, 32.66, 30.71, 30.47, 30.34, 30.32, 30.25, 29.93, 29.74, 28.16, 27.08, 26.52, 26.40, 23.63, 21.49, 14.66, 14.44.

[0374] C22H48N4O MS (ESI+) m / z=385.3845 (100%,[M+H]+).

[0375] PAFA26 has the following chemical formula:

[0376]

[0377] 1H NMR (400MHz, MeOD) δ: 5.38 (m, 14H), 3.23 (t, J = 6.83Hz, 3H), 2.86 (dd, J = 6.64, 12.11Hz, 12H), 2.66 (m, 8H), 2.40 (m, 3H), 2.24 (m, 3H), 2.10 (m, 3H), 1.71 (dq, J = 7.41, 14.31Hz, 6H), 0.99 (t, J = 7.55Hz, 3H).

[0378] 13C NMR (101MHz, MeOD) δ: 175.46, 132 .81, 130.18, 129.46, 129.25, 129.24, 129.21, 129.19, 129.14, 129.09, 128.96, 128.91, 128.17, 48.74, 48.72, 48.21, 47.79, 40

[0379] C31H54N4O MS(ESI+) m / z = 499.4331 (100%, [M+H]+).

[0380] PAFA27 has the following chemical formula:

[0381]

[0382] 1H NMR (400MHz, MeOD) δ: 3.15 (dt, J = 6.63, 26.81Hz, 3H), 2.65 (m, 10H), 2.14 (t, J = 7.51Hz, 2H), 1.61 (m, 11H), 1.27 (dd, J = 4.40, 8.38Hz, 14H), 0.8δ (m, 3H).

[0383] 13C NMR (101MHz, MeOD) δ: 176.35, 166.02, 50.23, 50.21, 49 .88, 48.05, 47.58, 47.50, 46.59, 40.40, 39.33, 37.94, 37.14, 33.04, 31.92, 30.62, 30.47, 30.41, 30.33, 30.18, 29.97, 29.88, 27.97, 27.89, 27.61, 27.10, 26.86, 23.73, 14.49.

[0384] C20H44N4O MS (ESI+) m / z=357.3549 (100%,[M+H]+).

[0385] They were all tested in combination with 2 μg / mL doxycycline. Specification 32 / 33 pages 41 CN 121443283 A

[0386] Figure 1 reports the antibacterial activity (MIC, in μg / mL) of doxycycline alone, compound (I) as defined in this invention alone, and combination of doxycycline with said compound (I) against bacterial strain (PA01) of Pseudomonas aeruginosa. More specifically, Figure 1 shows the concentrations (μg / mL) of PAFA6, PAFA13 to PAFA19 required to restore the activity of doxycycline against Pseudomonas aeruginosa PA01 at 2 μg / mL.

[0387] Figure 2 reports the antibacterial activity (MIC, in μg / mL) of compound (I) as defined in this invention alone, and combination of doxycycline with said compound (I) against bacterial strain (PA01) of Pseudomonas aeruginosa. More specifically, Figure 2 shows A) the MIC (μg / mL) of derivatives PAFA2, PAFA3, PAFA5, PAFA6, PAFA10, PAFA13 to PAFA27 against Pseudomonas aeruginosa PA01; and B) the concentrations (μg / mL) of derivatives PAFA2, PAFA3, PAFA5, PAFA6, PAFA10, PAFA13 to PAFA27 required to restore the activity of doxycycline against Pseudomonas aeruginosa PA01 at 2 μg / mL.

[0388] Furthermore, it was demonstrated that all the polyamined fatty acid compounds prepared as above did not exhibit toxicity to mammalian cells (data not shown). Specification 33 / 33 pages 42 CN 121443283 A Figure 1 Specification Figure 1 / 2 pages 43 CN 121443283 A Figure 2 Specification Figure 2 / 2 pages 44 CN 121443283 A,

Claims

1. Use of a polyaminated fatty acid compound, a pharmaceutically acceptable salt thereof and / or a solvate thereof corresponding to the following formula (I) as an adjuvant for an antibacterial agent: wherein: - m is an integer from 2 to 10; - n represents, independently in each occurrence of p, an integer from 1 to 5; and - p is an integer from 0 to 4. R 1 -C(=O)-NH-(CR 2 R 3 ) m -[X-(CR 4 R 5 ) n ] p -NR 6 R 7 (I) 5. Use according to any one of the preceding claims, wherein p is from 1 to 3. -R 1 represents a linear aliphatic chain comprising at least 7 carbon atoms, wherein the linear aliphatic chain is a mono-unsaturated aliphatic chain or a poly-unsaturated aliphatic chain, or a saturated aliphatic chain; - R 2 and R 3 each independently of one another, in each occurrence of m, independently represent a hydrogen atom or a Ci-C8alkyl group; - R 4 and R 5 independently of each other, independently in each occurrence of n, independently in each occurrence of p, represents a hydrogen atom or a Ci-C8alkyl group; - X represents, independently in each occurrence of p, -NR 8 - a group or a divalent 5- to 7-membered heterocycloalkyl comprising at least one nitrogen atom; wherein R 8 represents a hydrogen atom, a C1-C6 alkyl group, or -(CH2) q -NH2, wherein q represents an integer from 1 to 5; -R 6 and R 7 independently of one another represent a hydrogen atom, a Ci-C8alkyl group, or R 6 and R 7 form, together with the nitrogen atom to which they are attached, a 5- to 7- membered heterocyclyl group optionally substituted by one to three R 9 representing 9 a hydrogen atom, a halogen atom, a Ci-C8alkyl group, a C3-C7cycloalkyl group, a C1-C8haloalkyl group, a C1-C8alkoxy group, a C3-C7cycloalkoxy group, a C1-C8haloalkoxy group, a C6-C10aryl group, a C6-C10aryloxy group, a C -(CH2) u -NH2, wherein u represents an integer from 1 to 5; 8. Use according to any one of the preceding claims, wherein the polyaminated fatty acid compound is selected from the following compounds, a pharmaceutically acceptable salt thereof and / or a solvate thereof:

9. A pharmaceutical composition comprising at least one polyaminated fatty acid compound as defined in any one of the preceding claims and at least one antibacterial agent.

10. The pharmaceutical composition according to claim 9, wherein the pharmaceutical composition comprises from 0.1 to 50% by weight of the polyaminated fatty acid compound, based on the total weight of the pharmaceutical composition.

2. Use according to claim 1, wherein the straight-chain aliphatic chain R 1 is a monounsaturated aliphatic chain or a polyunsaturated aliphatic chain.

3. Use according to claim 1 or claim 2, wherein R 2 and R 3 independently of each other, in each occurrence of m, independently represent a hydrogen atom or a C1-C3 alkyl group.

4. Use according to any one of the preceding claims, wherein R 4 and R 5 independently of each other, independently in each occurrence of n, independently in each occurrence of p, represent a hydrogen atom or a C1-C3 alkyl group.

11. The pharmaceutical composition according to claim 9 or claim 10, wherein the weight ratio polyaminated fatty acid compound / antibacterial agent is from 0.2 to 2.

6. Use according to any one of the preceding claims, wherein X represents independently in each occurrence of p times -NR 8 a group, Ci-C3alkyl or -(CH2)q-NH2, wherein R 8 represents a hydrogen atom, wherein q represents an integer from 1 to 3.

7. Use according to any one of the preceding claims, wherein R 6 and R 7 are hydrogen atoms.

12. The pharmaceutical composition according to any one of claims 9 to 11, wherein the antibacterial agent is selected from the group consisting of tetracyclines and macrolides.

13. A pharmaceutical composition according to any one of claims 9 to 12 for use as a medicament.

14. The pharmaceutical composition for use according to claim 13, for use in the treatment of a bacterial disease; in particular a bacterial disease caused by Gram-negative bacteria.

15. The pharmaceutical composition for use according to claim 13, for use in killing or inhibiting the growth of persister cells.

16. A kit comprising: - a pharmaceutical composition comprising a polyaminated fatty acid compound as defined in any one of claims 1 to 8, and - a separate pharmaceutical composition comprising at least one antibacterial agent.

17. A polyaminated fatty acid compound selected from the following compounds of formula (I'-a): a pharmaceutically acceptable salt thereof and / or a solvate thereof. ​ ​ ​ ​ ​ ​ ​

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