Mono-substituted ester porphyrin derivative, preparation method thereof and application of mono-substituted ester porphyrin derivative as photosensitizer

Abstract

The present invention relates to a compound of formula (I) or a salt thereof, wherein the group R is as defined in the description; the invention also relates to a preparation method of the compound and application of the compound as a photosensitizer in photodynamic therapy. The mono-substituted protoporphyrin derivative has high permeability, high activity and good biological safety, the preparation method is simple in process and low in cost, a target product can be obtained with good yield, and the mono-substituted protoporphyrin derivative is especially suitable for industrial production and has wide application prospects. The compound can be used as a high-efficiency photosensitizer for treating oral diseases # imgabs0 #, such as periodontitis, gingivitis, dental plaque, decayed teeth and the like, caused by bacteria such as Gram-negative anaerobic bacillus, such as porphyromonas gingivalis in photodynamic therapy.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to a monosubstituted protoporphyrin derivative, a preparation method thereof and use thereof as a photosensitizer, and more specifically relates to a monosubstituted ester porphyrin derivative, a preparation method thereof and use thereof as a photosensitizer in photodynamic therapy. Background Art

[0002] Photodynamic therapy (PDT) is a new type of therapy that uses photosensitizers, light, and oxygen molecules to produce photodynamic reactions, and then selectively targets diseases such as malignant tumors, vascular lesions, and microbial infections. Photosensitizers are the core of PDT. The photosensitizers currently used in clinical practice mainly include Photofrin from the United States, Photogem from Russia, and Photosan from Germany. However, these photosensitizers still have many disadvantages. For example, they are mostly mixed preparations composed of porphyrin derivatives, the active ingredients are unclear, and there are no controllable quality standards. In addition, they can easily cause skin phototoxic effects such as rashes and blisters because they can remain in the skin for several weeks. Patients need to avoid direct sunlight for 1 month or even longer after taking the medicine.

[0003] 5-aminolevulinic acid (5-ALA) is a second-generation photosensitizer developed in recent years. It itself has no photosensitivity. After exogenous 5-ALA enters the body, it can be selectively absorbed and accumulated by actively proliferating cells and converted into protoporphyrin IX (PPIX) in the cells. PPIX in the cells is a photosensitive substance. After being irradiated with red light of a specific wavelength, a photodynamic reaction occurs, generating reactive oxygen species such as singlet oxygen, which kills actively proliferating cells.

[0004] Periodontitis is a chronic inflammatory disease that is mainly caused by the destruction of periodontal tissues by bacteria in dental plaque. Porphyromonas gingivalis is a Gram-negative coccobacillus and the main pathogen of periodontitis and gingivitis. Currently, drugs such as metronidazole tablets and amoxicillin capsules can be used for treatment, but frequent use of antibiotics will produce drug resistance, and some patients may even develop adverse reactions such as allergies, so safer, less toxic, and non-invasive alternative therapies are needed.

[0005] In summary, the development of photosensitizers with high antibacterial ability is of great significance. Summary of the invention

[0006] The present invention aims to provide a class of monosubstituted ester porphyrin derivatives, a preparation method thereof and use thereof as a photosensitizer, and more specifically to use them for inhibiting bacteria in photodynamic therapy to treat diseases such as periodontitis, gingivitis, dental plaque or caries.

[0007] The first aspect of the present invention relates to a compound of formula (I) or a salt thereof:

[0008] in: R stands for -OCH(R 1 )(R 2 ), R 1 represents hydrogen or (C1-C8)-alkyl, R 2 represents a 3- to 10-membered heterocyclic group, wherein the heterocyclic group contains one or more heteroatoms selected from O, S and N.

[0009] The second aspect of the present invention provides a method for preparing a compound of formula (I) or a salt thereof, the method comprising subjecting protoporphyrin (PPIX) to a condensation reaction with a compound having a reactive hydrogen atom in a polar organic solvent in the presence of a catalyst, a condensation agent and a condensation activator, wherein the compound having a reactive hydrogen atom is selected from a compound of general formula (II): RH (II) wherein R is as defined above.

[0010] The third aspect of the present invention relates to use of the compound or salt thereof according to the first aspect of the present invention as a photosensitizer for photodynamic therapy or use in the preparation of a photosensitizer for photodynamic therapy.

[0011] The fourth aspect of the present invention relates to use of a compound or a salt thereof according to the first aspect of the present invention in the preparation of a medicament for treating a disease or condition caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli.

[0012] The present invention also relates to a method for inhibiting Gram-negative or Gram-positive cocci, bacilli or coccobacilli for non-therapeutic purposes, the method comprising contacting the compound or its salt with Gram-negative or Gram-positive cocci, bacilli or coccobacilli and irradiating the compound or its salt with a specific wavelength of light having an effective inhibitory amount. The contact can be carried out in vivo or in vitro, for example, in vitro.

[0013] The present invention also relates to a method for treating a disease or condition caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli, which comprises administering a therapeutically effective amount of the compound or a salt thereof to a subject in need thereof and irradiating a therapeutically effective amount of light of a specific wavelength, preferably, wherein the subject is a mammal, more preferably a human.

[0014] The fourth aspect of the present invention relates to a pharmaceutical composition comprising the compound or salt thereof according to the first aspect of the present invention, and optionally further comprising one or more other active compounds.

[0015] Surprisingly, the present invention has the following beneficial effects: The present invention develops a novel class of monosubstituted ester porphyrin derivatives, which can be used as photosensitizers for photodynamic therapy. The compounds of the present invention all have high permeability, high activity and good biosafety, and the preparation method thereof is simple, low-cost, can obtain the target product in good yield and is particularly suitable for industrial production. It can be used as a high-efficiency photosensitizer in photodynamic therapy, has good antibacterial effect on bacteria such as Porphyromonas gingivalis, and has a therapeutic effect in oral diseases such as periodontitis, gingivitis, dental plaque and caries. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is the ultraviolet absorption spectrum of compound D-01 of the present invention.

[0017] Figure 2 The optical density (OD) of each test group in the antibacterial experiment and the antibacterial rate against Porphyromonas gingivalis are shown (compared with the blank light group in ac in the figure, compared with the PPIX group in d in the figure, **** P <0.0001). DETAILED DESCRIPTION

[0018] In order to better understand the present invention, the present invention will be described in detail below in conjunction with the embodiments and drawings. However, it should be understood that these embodiments and drawings are only for illustrative purposes only and are not intended to limit the present invention.

[0019] definition Unless otherwise specified, the terms "include" and "comprising" mentioned in this application may be open-ended or closed-ended. For example, the terms "include" and "comprising" may mean that other members, elements or method steps not listed may also be included or comprised, or may only mean that other members, elements or method steps listed may be included or comprised.

[0020] As used herein, the term "optionally" means that the subsequently described event, circumstance or material may or may not occur or exist, and that such description includes instances in which the event, circumstance or material occurs or exists and instances in which the event, circumstance or material does not occur or exist.

[0021] Unless otherwise defined, in the case of a specified group, for example in -OCH(R 1 )(R 2 ), the connection to the backbone or the rest of the molecule is via the first mentioned structural element, i.e., for example in -OCH(R 1 )(R 2 ) are connected through oxygen atoms.

[0022] As used herein, the term "alkyl" refers to a saturated, straight-chain or branched hydrocarbon radical having in each case the specified number of carbon atoms, such as (C1-C8)-alkyl, (C1-C6)-alkyl and (C1-C4)-alkyl, examples of which include, but are not limited to, methyl, ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl, 2-methylpropyl, 1,1-dimethylethyl, pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl, 1-ethylpropyl, hexyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 4- methylpentyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1-ethylbutyl, 2-ethylbutyl, 1,1,2-trimethylpropyl, 1,2,2-trimethylpropyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, n-heptyl, 1-methylhexyl, 2-methylhexyl, 1,1-dimethylpentyl, 1-ethylpentyl, 2-ethylpentyl, 1-propylbutyl, n-octyl, 1-methylheptyl, 2-ethylhexyl, 1,3-dimethylhexyl and 1-ethyl-2-methylpentyl.

[0023] Unless otherwise defined, the term "heterocyclyl" means a saturated or partially saturated monocyclic ring of carbon atoms and at least one heteroatom in the ring. Preferably, the heterocyclyl is a 3-10 membered heterocyclyl, for example containing 2, 3, 4, 5 or 6 carbon atoms and 1 or 2 heteroatoms selected from oxygen, sulfur and nitrogen, which can be connected to the parent molecular moiety through any carbon atom or nitrogen atom contained in the heterocyclic ring. If the ring contains more than one oxygen atom, they are not directly adjacent. Examples of heterocyclyl include, but are not limited to, aziridine, oxirane; azetidine, oxetane, thietane; tetrahydrofuranyl, 1,3-dioxolane, tetrahydrothienyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, isoxazolidinyl, oxazolidinyl, thiazolidinyl, isothiazolidinyl; piperidinyl, hexahydropyridazinyl, hexahydropyrimidinyl, piperazinyl, tetrahydropyranyl, dioxanyl, tetrahydro Thiopyranyl, dithianyl, morpholinyl, 1,2-oxazepanyl, oxathianyl, thiomorpholinyl; oxepanyl, azepanyl, 1,4-diazepanyl, 1,4-oxazepanyl; dihydrofuranyl, 1,3-dioxolyl, dihydrothiophenyl, pyrrolinyl, dihydroimidazolyl, dihydropyrazolyl, dihydrooxazolyl, dihydrothiazolyl; pyranyl, thiopyranyl and thiazinyl.

[0024] As used herein, the expression "membered heterocycle" refers to a saturated or partially saturated hydrocarbon ring system having the specified number of ring atoms and at least one carbon atom in the ring is replaced by a heteroatom (e.g. selected from N, O and S), for example a ring system containing 3 to 10, more preferably 3 to 8, especially 5 or 6 ring atoms, and at least one nitrogen atom and up to three heteroatoms selected from N, O and S. If more than one oxygen atom is contained in the ring, they are not directly adjacent.

[0025] In the context of the present invention, reference to the salt of a compound of formula (I) means a pharmaceutically acceptable salt thereof, generally those salts that are considered safe and suitable for use in pharmaceutical preparations. Unless otherwise expressly stated, the term "pharmaceutically acceptable salt" refers to pharmaceutically acceptable acid and base addition salts and solvates. Such pharmaceutically acceptable salts include, but are not limited to, salts of the following acids: for example, hydrochloric acid, trifluoroacetic acid, phosphoric acid, hydrobromic acid, sulfuric acid, sulfinic acid, formic acid, p-toluenesulfonic acid, methanesulfonic acid, benzoic acid, citric acid, acetic acid, etc., preferably acetate. Non-toxic pharmaceutical base addition salts include bases: salts such as sodium, potassium, calcium, ammonium, etc. Those skilled in the art will recognize a variety of non-toxic pharmaceutically acceptable addition salts.

[0026] As used herein, the term "therapeutically effective amount" or "effective amount" or "effective dose" refers to the amount of an active compound or agent that elicits the biological or medicinal response that is sought or desired by a researcher, physician or other clinician in a tissue, system, animal, individual or human.

[0027] Administration and Dosage Compounds or medicines of the present invention can work systemically and / or locally. For this purpose, they can be administered with suitable routes of administration, for example, by oral, lung, nose, sublingual, tongue, cheek, rectum, vagina, dermal, transdermal, conjunctival or ear canal route administration, or by subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial injection or infusion, or as implant or stent administration. For example, oral, spray inhalation, rectal administration, nasal administration, vaginal administration, topical administration, parenteral administration such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial injection or input, or by a kind of explanted reservoir medication.

[0028] Oral mucosal administration, oral administration, intramuscular injection, local administration, intraperitoneal administration or intravenous administration are preferred. Oral mucosal administration includes local injection, local coating, local washing, oral gargle, local controlled / slow release, microneedle and other device administration.

[0029] The drug delivery system of the present invention can be a targeted drug delivery system, a controlled drug delivery system, and a regulated drug delivery system, or it can be a new delivery system, such as an emulsified drug delivery system, a bionic drug delivery system, a living cell delivery system, an exosome drug delivery system, a microneedle drug delivery system, a nano drug delivery system, or a protein or polypeptide delivery system.

[0030] For these administration routes, the compounds according to the invention can be administered in suitable administration forms.

[0031] The drug of the present invention can be administered in a unit dosage form. The dosage form can be a liquid dosage form, a semisolid preparation and a solid dosage form. The liquid dosage form can be a true solution, a colloid, a microparticle dosage form, a suspension dosage form, etc. The semisolid dosage form can be an ointment, a cream, a paste, a gel, etc. The solid dosage form can be an orodispersible film, a tablet, a capsule, a dripping pill, a pill, a powder, a granule, a suppository, a freeze-dried powder injection, an inclusion compound, an implant, a patch, etc.

[0032] The compounds according to the invention can be incorporated into the administration forms mentioned. This can be achieved in a known manner by mixing with pharmaceutically suitable carriers, excipients and / or other auxiliaries.

[0033] The single administration dose of the medicament of the present invention is 0.01-100 mg active ingredient / kg body weight, preferably 0.02-80 mg active ingredient / kg body weight, more preferably 0.05-50 mg active ingredient / kg body weight, still more preferably 0.08-40 mg active ingredient / kg body weight, particularly preferably 0.1-20 mg active ingredient / kg body weight, still preferably 0.1-15 mg active ingredient / kg body weight, for example 0.1-10 mg active ingredient / kg body weight, 0.1-8 mg active ingredient / kg body weight, most preferably 0.2-6 mg active ingredient / kg body weight.

[0034] In one embodiment, the medicament of the present invention is administered at least once a month, for example 1, 2, 3, 4 or 5 times a month. Preferably, the medicament of the present invention is administered 1, 2 or 3 times a month. Here, the term "active ingredient" refers to a compound of formula (I) or a salt thereof in the present invention.

[0035] plan The first aspect of the present invention relates to a compound of formula (I) or a salt thereof:

[0036] in: R stands for -OCH(R 1 )(R 2 ), R 1 represents hydrogen or (C1-C8)-alkyl, R 2 represents a 3- to 10-membered heterocyclic group, wherein the heterocyclic group contains one or more heteroatoms selected from O, S and N.

[0037] In a preferred embodiment, R1 represents hydrogen and R2 represents a 3-10 membered heterocyclic group, wherein the heterocyclic group contains one or more heteroatoms selected from O and N.

[0038] In a preferred embodiment, R 1 represents hydrogen or (C1-C6)-alkyl, R 2 represents a 3-8 membered heterocyclic group, wherein the heterocyclic group contains 1 to 3 heteroatoms selected from O, S and N. Further preferably, R 1 Represents hydrogen, R 2 represents a 3- to 8-membered heterocyclic group, wherein the heterocyclic group contains 1 to 3 heteroatoms selected from O and N.

[0039] In a preferred embodiment, the 3-8 membered heterocyclic group is a 3-8 membered heterocyclic group containing 1-3 (preferably 1-2) O atoms, 1-3 (preferably 1-2) N atoms and / or 1-3 (preferably 1-2) S atoms.

[0040] In a further preferred embodiment, R 1 Represents hydrogen, R 2 represents a 5- to 7-membered heterocyclic group, wherein the heterocyclic group contains 1 to 3 heteroatoms selected from O and N.

[0041] In a further preferred embodiment, R 1 Represents hydrogen, R 2 It represents a morpholinyl group, preferably a morpholin-3-yl group.

[0042] The definitions of radicals listed above in general terms or in preferred ranges can be combined with one another as desired, ie including combinations between the preferred ranges given.

[0043] Very particularly preferred compounds according to the invention are .

[0044] The second aspect of the present invention provides a method for preparing a compound of formula (I) or a salt thereof, the method comprising:

[0045] In the presence of a catalyst, a condensation agent and a condensation activator, protoporphyrin (PPIX) is subjected to a condensation reaction with a compound having a reactive hydrogen atom in a polar organic solvent, wherein the compound having a reactive hydrogen atom is selected from the compounds of the general formula (II): RH (II) wherein R is as defined above.

[0046] The catalyst suitable for the method of the present invention is an organic base catalyst, which is preferably selected from N,N-diisopropylethylamine (DIPEA), N,N-dimethylcyclohexylamine, N,N-dimethylbenzylamine, N-methylmorpholine (NMM), N-ethylmorpholine, N-methylimidazole (NMI), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), triethylamine, pyridine, N,N-dimethylaminopyridine (DMAP), 2,6-lutidine or a mixture thereof, more preferably DIPEA, NMI, and particularly preferably DIPEA.

[0047] The condensing agent suitable for the method of the present invention is a carbodiimide type condensing agent, which is preferably selected from 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 1,3-dicyclohexylcarbodiimide (DCC), N,N-diisopropylcarbodiimide (DIC) or a mixture thereof, more preferably EDCI.

[0048] Examples of condensation activators suitable for the method of the present invention include, but are not limited to, N,N-dimethylaminopyridine (DMAP), 4-pyrrolidinylpyridine (4-PPY), 1-hydroxy-7-azabenzotriazole (HOAT), 1-hydroxybenzotriazole (HOBT), N-hydroxysuccinimide (NHS), N-hydroxyphthalimide (NHPI), pentafluorophenol (PFP), etc., preferably HOBT.

[0049] The polar organic solvent suitable for the method of the present invention may be preferably selected from N,N-dimethylformamide (DMF), N,N-dimethylacetamide, formamide, dimethyl sulfoxide (DMSO), acetone, pyridine or a mixture thereof, more preferably DMF.

[0050] In the method of the present invention, the molar ratio of the protoporphyrin to the catalyst is 1:(0.6-1.5), preferably 1:(0.7-1.3), and more preferably 1:(0.8-1.1).

[0051] In the method of the present invention, the molar ratio of the compound having reactive hydrogen atoms to the condensation agent or condensation activator is 1:(0.6-1.5), preferably 1:(0.7-1.3), and more preferably 1:(0.8-1.1).

[0052] In a preferred embodiment of the present invention, the compound having a reactive hydrogen atom is (R)-tert-butyl 3-(hydroxymethyl)morpholine-4-carboxylate.

[0053] According to the reaction requirements, the compound having a reactive hydrogen atom may optionally carry an amino protecting group commonly used in amidation reactions known to those skilled in the art, such as tert-butyloxycarbonyl (Boc). The removal of the amino protecting group may be carried out by conventional methods known to those skilled in the art, for example, using a solution of hydrogen chloride in a specific organic solvent (e.g., dioxane, ethyl acetate, methanol), preferably a solution of hydrogen chloride in dioxane.

[0054] The method for preparing the compound of formula (I) of the present invention uses a combination of a specific organic base catalyst, a condensation agent and a condensation activator, especially a combination of DIPEA, EDCI and HOBT in a specific ratio, and can obtain a target compound with high purity in a good yield through simple post-treatment. If the amount of the catalyst, condensation agent or condensation activator is too low or too high, it will have an adverse effect on the target product, such as difficulty in post-treatment, increased proportion of by-products, and low yield of the target product.

[0055] In the method of the present invention, the condensation reaction time is 0.5-15 hours, preferably 1-10 hours, more preferably 2-7 hours.

[0056] In the above method for preparing the compound of the present invention, the method may optionally include other post-treatment steps. The post-treatment steps may include conventional purification steps such as adjusting the pH value, crystallization, extraction, filtration, concentration under reduced pressure, and drying. Each of the above steps can be carried out in a conventional manner known to those skilled in the art. If present, extraction is usually carried out using a mixed solution of dichloromethane and methanol, preferably a mixed solution of dichloromethane / methanol = 10:1 (v / v); drying is usually carried out by freeze drying, infrared drying, vacuum drying, etc., preferably freeze drying.

[0057] In a preferred embodiment of the present invention, the method may further include a purification step by a chromatographic column. The purification may be performed by: a normal phase silica gel column, the silica gel particle size is 30-100 μm, preferably 40-63 μm, the loading is 20-120 g, preferably 40 g, eluted with dichloromethane / methanol containing 5% acetic acid (v / v) (elution gradient 100% / 0% to 90% / 10%, gradient elution time 15 min); or a reverse phase cyano column, the particle size is 10-100 μm, preferably 20-45 μm, the loading is 20-120 g, preferably 40 g, eluted with water containing 5% acetic acid / acetonitrile containing 5% acetic acid (v / v) (elution gradient 95% / 5% to 5% / 95%, gradient elution time 20 min).

[0058] The compounds of the present invention can be prepared according to the above methods. However, it should be understood that a person skilled in the art, based on his own general knowledge and available publications, can adjust the methods according to the specific circumstances of the respective compounds of the present invention that he wishes to synthesize.

[0059] The third aspect of the present invention relates to use of the compound or salt thereof according to the first aspect of the present invention as a photosensitizer for photodynamic therapy or use in the preparation of a photosensitizer for photodynamic therapy.

[0060] The fourth aspect of the present invention relates to use of a compound or a salt thereof according to the first aspect of the present invention in the preparation of a medicament for treating a disease or condition caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli.

[0061] The present invention also relates to a method for inhibiting Gram-negative or Gram-positive cocci, bacilli or coccobacilli, which comprises contacting the compound or its salt with Gram-negative or Gram-positive cocci, bacilli or coccobacilli and irradiating the compound with a specific wavelength of light having an effective inhibitory amount. Preferably, the inhibition is for non-therapeutic purposes. The contact can be carried out in vivo or in vitro, for example, in vitro.

[0062] The present invention also relates to a method for treating a disease or condition caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli, which comprises administering a therapeutically effective amount of the compound or a salt thereof to a subject in need thereof and irradiating a therapeutically effective amount of light of a specific wavelength, preferably, wherein the subject is a mammal, more preferably a human.

[0063] The compound or salt thereof according to the first aspect of the present invention is used as a photosensitizer in photodynamic therapy, specifically for treating diseases or conditions caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli.

[0064] The Gram-negative or -positive cocci, bacilli or coccobacilli can be selected from Gram-negative bacilli or coccobacilli, or Gram-positive cocci; for example, Gram-negative anaerobic bacilli or coccobacilli, Gram-negative facultative anaerobic coccobacilli, Gram-negative aerobic bacilli or Gram-positive facultative anaerobic cocci.

[0065] In a preferred embodiment, the Gram-negative or Gram-positive cocci, bacilli or coccobacilli is Porphyromonas gingivalis ( Porphyromonas gingivalis ), Actinobacillus actinomycetemcomitans ( Actinobacillus actinomycetemcomitans ), Tannerella forsythiae ( Tannerella forsythia )、Fusobacterium nucleatum( Fusobacterium nucleatum ), Prevotella intermedia ( Prevotella intermedia ), Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) or Staphylococcus aureus ( Staphylococcus aureus gingivalis); preferably Porphyromonas gingivalis.

[0066] In a preferred embodiment, the disease or condition is selected from one or more of the following: periodontitis, gingivitis, dental plaque and caries; preferably, the periodontitis is chronic periodontitis or aggressive periodontitis; Preferably, the disease or condition is selected from one or more of the following: swollen gums, bleeding gums, painful gums, bad breath, periodontal pocket formation, alveolar bone resorption and loose teeth.

[0067] In a preferred embodiment, the compound of formula (I) or a salt thereof is used as a photosensitizer in photodynamic therapy to treat diseases or conditions caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli.

[0068] Preferably, the irradiation time of light irradiation is 10 s-2000 s, preferably 20-1500 s, further preferably 30-1200 s, more preferably 60-900 s; for example, 20 s, 30 s, 60 s, 120 s, 240 s, 480 s, 600 s, 900 s, 1200 s, 1500 s, etc.

[0069] Preferably, the light dose of light irradiation is 1-300 J, preferably 5-200 J, more preferably 10 J-180 J, more preferably 20-120 J; for example, 10 J, 20 J, 30 J, 40 J, 60 J, 80 J, 100 J, 120 J, 150 J, 180 J, etc.

[0070] Preferably, the wavelength range of light irradiation is 300 nm-800 nm, preferably 350 nm-700 nm, more preferably 400 nm-650 nm, for example, 405 nm, 505 nm, 540 nm, 575 nm or 630 nm; preferably 405 nm.

[0071] Preferably, the optical power of the light irradiation is 30 mW-1000 mW, preferably 60 mW-600 mW, more preferably 80 mW-300 mW.

[0072] In the context of the present invention, the term "light dose" refers to the actual light dose at the light irradiation site, which is obtained by multiplying the light power by the irradiation time.

[0073] In the context of the present invention, the term "optical power" refers to the actual optical power at the light irradiation site, which is measured by an optical power meter (model: PM100D) purchased from THORLABS to determine the actual power at the treatment site.

[0074] Preferably, in the context of the present invention, the salt is a pharmaceutically acceptable salt.

[0075] The fifth aspect of the present invention relates to a pharmaceutical composition comprising a compound or a salt thereof according to the first aspect of the present invention, and optionally further comprising one or more other active compounds. The other active compounds are compounds that are generally effective for treating diseases or conditions caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli.

[0076] In one embodiment, the pharmaceutical composition is a kit, which further comprises instructions for using the compound or salt thereof as a photosensitizer in photodynamic therapy.

[0077] In a preferred embodiment, the drug comprises 1 mg-600 mg, preferably 1 mg-400 mg, more preferably 2 mg-300 mg, further preferably 3 mg-200 mg, further preferably 4 mg-100 mg of a compound of formula (I) or a salt thereof, for example 2 mg-500 mg, 2 mg-400 mg, 2 mg-300 mg, 3 mg-240 mg, 5 mg-120 mg or 5 mg-60 mg, more specifically for example 2 mg, 4 mg, 10 mg, 12 mg, 15 mg, 18 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 80 mg, 100 mg, 120 mg, 180 mg, 240 mg, 300 mg, 400 mg or 600 mg.

[0078] In a preferred embodiment, the medicine also includes a pharmaceutically acceptable carrier, excipient and / or other adjuvants. When comprising a pharmaceutically acceptable carrier, excipient and / or other adjuvants, usually an effective dose of the compound of formula (I) or its salt and one or more pharmaceutically acceptable carriers, excipients and / or other adjuvants are combined to form a suitable form of administration or dosage form, and this procedure includes mixing, granulating, compressing, dissolving or lyophilizing the components by a suitable method. The content of the carrier in the medicine can be 1 to 98 weight %, usually accounting for about 80 weight %. For convenience, other adjuvants such as local anesthetics, preservatives, buffers can be directly dissolved in the carrier.

[0079] Example Detailed synthesis examples of selected compounds of the present invention are given below. However, these examples are merely illustrative and should not be interpreted as limiting the scope of the present invention in any way.

[0080] The NMR peaks reported in the synthetic examples are 1H NMR spectral data were obtained on a Bruker 400 MHz nuclear magnetic resonance spectrometer and the signals listed have the meanings given below: s = singlet, d = doublet, m = multiplet. The deuterated solvents used in each case are also specified in the table.

[0081] In the present invention, in addition to the above-mentioned NMR peak list form 1 In addition to H NMR spectral data, the structures of the compounds prepared in the synthetic examples were also characterized by liquid chromatography-mass spectrometry (LC-MS). In the present invention, the liquid chromatography-mass spectrometry (LC-MS) used to detect mass spectrometry data was purchased from Waters Technology Shanghai Co., Ltd., model: SQD2; the medium-pressure column machine used for compound separation and purification was purchased from Santai Technology (Changzhou) Co., Ltd., model: SepaBean mechine T.

[0082] Unless otherwise explained or defined, scientific and technical terms used in connection with the present invention shall have the meanings that are commonly understood by one of ordinary skill in the art.

[0083] The reagents or instruments used without indicating the manufacturer are all commercially available conventional products commonly used in the art. In the present invention, unless otherwise specified, all operations are carried out at room temperature and normal pressure. Unless otherwise specified, the contents and percentages in the context of this application are based on weight.

[0084] A. Synthesis Examples Example 1: Synthesis of D-01 Protoporphyrin (200 mg, 0.36 mmol, 1 equ) was dissolved in 10 mL DMF, and EDCI (61 mg, 0.32 mmol, 0.9 equ), HOBt (43 mg, 0.32 mmol, 0.9 equ) and DIPEA (41 mg, 0.32 mmol, 0.9 equ) were added. The mixture was stirred at room temperature for 10 min, and (R)-tert-butyl 3-(hydroxymethyl)morpholine-4-carboxylate (78 mg, 0.36 mmol, 1.0 equ) was added. After stirring at room temperature for 4 h, the reaction solution was poured into 30 mL of water containing 10% citric acid, and extracted three times with a mixed solution of dichloromethane / methanol (10 / 1, v / v) (30 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by normal phase silica gel column (dichloromethane / methanol containing 5% acetic acid (v / v) 100% / 0% to 90% / 10%, elution time 15 min), and after evaporating the solvent, lyophilized to obtain 149 mg of intermediate D-01-01 with a yield of 55%.

[0085] At 0°C, 149 mg of intermediate D-01-01 was dissolved in 5 mL of 4 mol / L hydrogen chloride dioxane solution and stirred at this temperature for 2 h. After the reaction was completed by liquid chromatography-mass spectrometry, the product was concentrated under reduced pressure at room temperature and lyophilized to obtain product D-01 (124 mg, yield 95%).

[0086] LC-MS(m / z): 662.6 [M+H] + .

[0087] 1 H NMR (400 MHz, DMSO- d 6): δ 10.28–9.77 (m, 4H), 8.52–8.17 (m, 2H), 6.39 (d, J = 16.0 Hz, 2H), 6.19 (d, J = 9.6 Hz, 2H), 4.24 (s, 4H), 4.15 (m, 2H), 3.97 (m, 2H), 3.61 (s, 12H), 3.11–2.96 (m, 5H), 2.73 (s, 4H).

[0088] B. Absorption spectra of the compounds of the present invention In order to explore the optimal excitation wavelength of the drug, the absorption wavelength of the compound of the present invention is detected. The specific steps are as follows: A dimethyl sulfoxide solution (brand: Shanghai Runjie) of the porphyrin derivative D-01 of the present invention was prepared at a concentration of 20 μg / mL, 4 mL each, and placed in a cuvette. An ultraviolet spectrophotometer (purchased from Shanghai Yuanxi Instrument Co., Ltd., model: X-8S) was used to test the absorption spectrum of the solution in the cuvette.

[0089] like Figure 1 As shown, the compound of the present invention can absorb energy at 405, 505, 540, 575 and 630 nm, especially the strongest absorption energy near 405 nm. According to the above results, the excitation wavelength of the porphyrin derivative of the present invention is determined to be 405 nm and 630 nm. Among them, the peak shape of protoporphyrin is not smooth, indicating that the molecule has molecular aggregation at this concentration. At the same time, it is suggested that the ACQ effect of the compound of the present invention is weaker than that of protoporphyrin, and it is less likely to occur intermolecular aggregation at the same concentration, and the PDT effect is stronger.

[0090] C. Effect Example Example 1 1. Materials and Methods 1.1 Information on the drug to be tested Test drug: Compound D-01 of the present invention prepared in Synthesis Example 1.

[0091] Positive drug 1: protoporphyrin (PPIX).

[0092] Positive drug 2: 5-aminolevulinic acid (5-ALA).

[0093] Solvent: DMSO.

[0094] 1.2 Experimental methods Porphyromonas gingivalis (purchased from Beijing Beina Chuanglian Biotechnology Research Institute, batch number BNCC353909) was revived and inoculated into BHI medium (brain heart infusion medium) and cultured to the logarithmic phase under anaerobic conditions of 10% H2, 10% CO2, 80% N2, and 37°C; 1 mL of bacterial solution was taken, the OD value was measured, and after centrifugation and discarding the supernatant, a certain amount of BHI medium was added to adjust the bacterial solution concentration to 10 10 CFU / mL (1 OD≈9.52*10 9 CFU / mL) for later use.

[0095] Test drug group: add 100 μL bacterial solution, 898 μL BHI medium and 2 μL DMSO solution of the drug to be tested to the centrifuge tubes respectively, shake well (the final concentration of bacterial solution is 10 9 CFU / mL, the final concentration of the test drug was 10 μg / mL, the final concentration of positive drug 1 was 10 μg / mL, and the final concentration of positive drug 2 was 80 μg / mL). Protect from light, culture under the same anaerobic conditions for 4 h, centrifuge and discard the supernatant, then resuspend with 1 mL of BHI medium, add the resuspended liquid to a 96-well plate, add 100 μL of bacterial solution to each well, repeat 3 times for each group, and set 4 replicates each time. Illuminate under a 405 nm laser, the laser output power is 300 mW, the irradiation is 66.67 s, and the light dose is 20 J. After the illumination, continue to culture under the same anaerobic conditions for 24 h, then measure the OD value of each group of bacterial solution at 600 nm and calculate the inhibition rate.

[0096] Blank illumination group (referred to as blank illumination group): the above method was followed, except that 100 μL of bacterial solution and 900 μL of BHI culture medium were added to the centrifuge tube.

[0097] Blank no-light group (referred to as blank group): the above method was followed, except that 100 μL of bacterial solution and 900 μL of BHI medium were added to the centrifuge tube without light treatment.

[0098] Vehicle group: The above method was followed, except that 100 μL of bacterial solution, 898 μL of BHI medium and 2 μL of DMSO were added to the centrifuge tube.

[0099] The antibacterial rate calculation formula is as follows:

[0100] Among them: OD 阴性 It represents the average OD value of bacterial solution measured in the blank illumination group; OD 待测 It represents the average OD value of bacterial solution measured in the test drug group.

[0101] 1.3 Calculation of statistical differences GraphPad Prism8 was used for statistical analysis and OD 600 The antibacterial effect was evaluated by the difference in values. One-way ANOVA was used for multiple group comparisons, and post hoc multiple comparisons were performed. P<0.05 was considered statistically significant, among which *<0.05, **<0.01, *** ＜ 0.001, ****<0.0001, represents a significant decrease.

[0102] 2. Results and Discussion 2.1 Results of the first biological replication The results are as follows Figure 2 As shown in (a), compared with the blank illumination group, the optical density of compound D-01 and the optical density of positive drug 1 were significantly reduced (p<0.0001), and their inhibition rates were 64.14% and 32.65%, respectively.

[0103] 2.2 Results of the second biological replication The results are as follows Figure 2 As shown in (b), compared with the blank illumination group, the optical density of compound D-01 and positive drug 1 was significantly reduced (p<0.0001). The inhibition rates of D-01 and positive drug 1 were 72.5% and 28.2%, respectively.

[0104] 2.3 Results of the third biological replication The results are as follows Figure 2 As shown in (c), compared with the blank illumination group, the optical density of compound D-01 and positive drug 1 was significantly reduced (p<0.0001). The inhibition rates of D-01 and positive drug 1 were 68.2% and 32.4%, respectively.

[0105] 2.4 Inhibition rate of three biological replicates The results are as follows Figure 2 As shown in (d) and Table 1, the compound D-01 of the present invention has a significant and relatively stable antibacterial effect, and the effect is significantly better than that of the control compounds PPIX and 5-ALA, with a significant difference (p<0.0001).

[0106] Table 1: Inhibitory effects of the compounds D-01, PPIX and 5-ALA on Porphyromonas gingivalis .

Claims

1. A compound of formula (I) or a salt thereof: (I) in: R means -OCH(R 1 )(R 2 ), R 1 represents hydrogen, R 2 represents a 3- to 10-membered heterocyclic group, wherein the heterocyclic group contains one or more heteroatoms selected from O and N.

2. The compound or salt thereof according to claim 1, wherein: R 1 represents hydrogen, R 2 represents a 3- to 8-membered heterocyclic group, wherein the heterocyclic group contains 1 to 3 heteroatoms selected from O and N.

3. The compound or salt thereof according to claim 2, wherein R 1 represents hydrogen, R 2 represents a 5- to 7-membered heterocyclic group, wherein the heterocyclic group contains 1 to 3 heteroatoms selected from O and N.

4. A method for preparing a compound of formula (I) or a salt thereof as claimed in any one of claims 1 to 3, comprising subjecting protoporphyrin to a condensation reaction with a compound having a reactive hydrogen atom in a polar organic solvent in the presence of a catalyst, a condensation agent and a condensation activator, wherein the compound having a reactive hydrogen atom is selected from a compound of general formula (II): RH (II) wherein R is as defined in any one of claims 1 to 3.

5. The method according to claim 4, wherein: The catalyst is an organic base catalyst selected from N,N-diisopropylethylamine, N,N-dimethylcyclohexylamine, N,N-dimethylbenzylamine, N-methylmorpholine, N-ethylmorpholine, N-methylimidazole, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, pyridine, N,N-dimethylaminopyridine, 2,6-lutidine or a mixture thereof; The condensing agent is a carbodiimide type condensing agent selected from 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1,3-dicyclohexylcarbodiimide, N,N-diisopropylcarbodiimide or a mixture thereof; The condensation activator is selected from N,N-dimethylaminopyridine, 4-pyrrolidinylpyridine, 1-hydroxy-7-azabenzotriazole, 1-hydroxybenzotriazole, N-hydroxysuccinimide, N-hydroxyphthalimide, pentafluorophenol; or The molar ratio of the compound having reactive hydrogen atoms to the condensation agent or condensation activator is 1:(0.6-1.5).

6. Use of a compound of formula (I) or a salt thereof according to any one of claims 1 to 3 in the preparation of a photosensitizer for photodynamic therapy.

7. Use of a compound of formula (I) or a salt thereof according to any one of claims 1 to 3 for the preparation of a medicament for the treatment of a disease or condition caused by a Gram-negative or Gram-positive coccus, bacillus or coccobacillus.

8. The method according to claim 7, wherein the Gram-negative or Gram-positive cocci, bacilli or coccobacilli are Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Tannerella forsythiae, Fusobacterium nucleatum, Prevotella intermedia, Pseudomonas aeruginosa or Staphylococcus aureus.

9. The use according to claim 7, wherein the disease or condition is selected from one or more of the following: periodontitis, gingivitis, dental plaque and caries; or the disease or condition is selected from one or more of the following: swollen gums, bleeding gums, gum pain, bad breath, periodontal pocket formation, alveolar bone resorption and loose teeth.

10. A pharmaceutical composition comprising a compound or a salt thereof according to any one of claims 1 to 3, optionally further comprising one or more other active compounds.

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