Monosubstituted ester porphyrin derivatives, their preparation methods and uses as photosensitizers

Abstract

The present invention relates to a compound of formula (I) or a salt thereof, wherein the group R is as defined in the specification; and also relates to a method for preparing the compound and its use as a photosensitizer in photodynamic therapy. The monosubstituted protoporphyrin derivatives involved in the present invention all have high permeability, high activity, and good biosafety. The preparation method is simple, low-cost, and can obtain the target product in good yield. It is particularly suitable for industrial production and can be used as a highly effective photosensitizer in photodynamic therapy to treat oral diseases such as periodontitis, gingivitis, dental plaque, and caries caused by Gram-negative anaerobic cocci and other bacteria such as Porphyromonas gingivalis.

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and specifically relates to monosubstituted protoporphyrin derivatives, their preparation methods and their use as photosensitizers, and more specifically to monosubstituted ester porphyrin derivatives, their preparation methods and their use as photosensitizers in photodynamic therapy. Background Art

[0002] Photodynamic therapy (PDT) is a novel treatment that uses photosensitizers, light, and oxygen molecules to produce a photodynamic reaction, selectively targeting diseases such as malignant tumors, vascular lesions, and microbial infections. The photosensitizer is the core of PDT. Currently, the main photosensitizers used clinically include Photofrin from the United States, Photogem from Russia, and Photosan from Germany. However, these photosensitizers have numerous drawbacks. For example, they are often mixed preparations composed of porphyrin derivatives, with unclear active ingredients and uncontrolled quality standards. Furthermore, due to their long-term retention in the skin for several weeks, they can easily cause phototoxic effects such as rashes and blisters. Patients must avoid direct sunlight for one month or even longer after treatment.

[0003] 5-Aminolevulinic acid (5-ALA) is a recently developed second-generation photosensitizer. While not inherently photosensitizing, exogenous 5-ALA is selectively absorbed and accumulated by actively proliferating cells, where it is converted into protoporphyrin IX (PPIX). PPIX within cells is a photosensitizer that, upon exposure to red light of a specific wavelength, undergoes a photodynamic reaction, generating reactive oxygen species such as singlet oxygen, which kills actively proliferating cells.

[0004] Periodontitis is a chronic inflammatory disease primarily caused by the destruction of periodontal tissues by bacteria in dental plaque. Porphyromonas gingivalis, a Gram-negative coccobacillus, is the primary pathogenic bacteria that causes periodontitis and gingivitis. Currently, treatment options include metronidazole tablets and amoxicillin capsules. However, frequent use of antibiotics can lead to drug resistance, and some patients may even experience adverse reactions such as allergies. Therefore, safer, less toxic, and non-invasive alternative treatments are needed.

[0005] In summary, the research and 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 photosensitizers, and more specifically to use them for antibacterial treatment in photodynamic therapy for 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]

[0009] Wherein:

[0010] R represents -OCH(R 1 )(R 2 ),

[0011] R 1 represents hydrogen or (C1-C8)-alkyl,

[0012] R 2 represents a 3- to 10-membered heterocyclic group, wherein said heterocyclic group contains one or more heteroatoms selected from O, S, and N.

[0013] 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 condensing agent, and a condensation activator, wherein the compound having a reactive hydrogen atom is selected from compounds of general formula (II)

[0014] RH (II)

[0015] wherein R is as defined above.

[0016] The third aspect of the present invention relates to the use of a compound or a salt thereof according to the first aspect of the present invention as a photosensitizer for photodynamic therapy or in the preparation of a photosensitizer for photodynamic therapy.

[0017] The fourth aspect of the present invention relates to the 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.

[0018] 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 a salt thereof with Gram-negative or Gram-positive cocci, bacilli, or coccobacilli and irradiating with light of a specific wavelength in an inhibitory effective amount. The contacting can be carried out in vivo or in vitro, for example, in vitro.

[0019] 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 to a subject in need a therapeutically effective amount of the compound or a salt thereof and irradiating with light of a therapeutically effective amount of a specific wavelength, preferably, wherein the subject is a mammal, more preferably a human.

[0020] The fourth aspect of the present invention relates to a pharmaceutical composition, which comprises the compound according to the first aspect of the present invention or a salt thereof, and optionally further comprises one or more other active compounds.

[0021] Surprisingly, the present invention has the following beneficial effects:

[0022] The present invention has developed 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. Their preparation methods have simple processes, low costs, can obtain target products in good yields and are particularly suitable for industrial production. They can be used as highly efficient photosensitizers in photodynamic therapy and have good antibacterial effects on bacteria such as Porphyromonas gingivalis, and have therapeutic effects on oral diseases such as periodontitis, gingivitis, dental plaque and dental caries. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is the ultraviolet absorption spectrogram of compound D-01 of the present invention.

[0024] Figure 2 Shows the optical density (OD) of each test group in the antibacterial experiment and the antibacterial rate against Porphyromonas gingivalis (in a-c in the figure compared with the blank light irradiation group, in d in the figure compared with the PPIX group, **** P <0.0001). DETAILED DESCRIPTION OF THE EMBODIMENTS

[0025] To better understand the present invention, the present invention will be described in detail below in conjunction with examples and drawings. It should be understood, however, that these examples and drawings are only for illustrative purposes of the present invention and are not intended to limit the present invention.

[0026] definition

[0027] Unless otherwise specified, the terms "comprising" and "including" mentioned in this application mean open-ended, and can also be closed-ended. For example, the said "comprising" and "including" can mean that other members, elements or method steps not listed can also be included or contained, or can only include or contain the other members, elements or method steps listed.

[0028] The term "optionally" used herein means that the events, circumstances or substances described later may occur or exist, or may not occur or exist, and such description includes the cases where the events, circumstances or substances occur or exist and the cases where the events, circumstances or substances do not occur or exist.

[0029] Unless otherwise differently defined, in the case of specifying a group, for example, in -OCH(R 1 )(R 2) In the case of, the connection to the skeleton or the rest of the molecule is through the first-mentioned structural element, i.e., for example, in the case of -OCH(R 1 )(R 2 ), it is connected through an oxygen atom.

[0030] As used herein, the term "alkyl" refers to a saturated straight-chain or branched hydrocarbon group having a specified number of carbon atoms in each case, such as (C1-C8)-alkyl, (C1-C6)-alkyl, and (C1-C4)-alkyl. Examples 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.

[0031] Unless otherwise defined, the term "heterocyclyl" refers to 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 may be attached to the parent molecular moiety via 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, tetrahydrothiophenyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, isoxazolidinyl, oxazolidinyl, thiazolidinyl, isothiazolidinyl; piperidinyl, hexahydropyridazinyl, hexahydropyrimidinyl, piperazinyl, tetrahydropyranyl, dioxanyl, tetrahydro Thiaphanyl, 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.

[0032] 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 in which 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 comprising 3 to 10, more preferably 3 to 8, in particular 5 or 6 ring atoms, and at least one nitrogen atom and up to three heteroatoms selected from N, O and S. If the ring comprises more than one oxygen atom, they are not directly adjacent.

[0033] In the context of the present invention, reference to a salt of a compound of formula (I) means a pharmaceutically acceptable salt thereof, generally a salt that is considered pharmaceutically safe and suitable for use in pharmaceutical formulations. 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 acids such as 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, and the like, preferably acetate. Non-toxic pharmaceutical base addition salts include salts of bases such as sodium, potassium, calcium, ammonium, and the like. Those skilled in the art will recognize a variety of non-toxic pharmaceutically acceptable addition salts.

[0034] As used herein, the term "therapeutically effective amount" or "effective amount" or "effective dose" refers to that amount of an active compound or agent that elicits the biological or medicinal response that is being sought or desired by a researcher, physician, or other clinician in a tissue, system, animal, individual, or human.

[0035] Administration and dosage

[0036] The compounds or drugs of the present invention can act systemically and / or locally. For this purpose, they can be administered by suitable routes of administration, such as by oral, pulmonary, nasal, sublingual, lingual, buccal, rectal, vaginal, dermal, transdermal, conjunctival or otic canal routes, or by subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial injection or infusion, or as implants or stents. For example, oral administration, 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 infusion, or administration with the aid of an implanted reservoir.

[0037] Oral mucosal administration, oral administration, intramuscular injection, topical administration, intraperitoneal or intravenous administration are preferred. Oral mucosal administration includes administration by devices such as local injection, topical coating, topical irrigation, oral gargling, local controlled / sustained release, and microneedles.

[0038] 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 can also be a novel delivery system, such as an emulsified delivery system, a biomimetic drug delivery system, a live 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, etc.

[0039] For these routes of administration, the compounds of the present invention can be administered in a suitable dosage form.

[0040] The drugs of the present invention can be administered in unit dosage forms. The dosage forms can be liquid dosage forms, semi-solid preparations and solid dosage forms. Liquid dosage forms can be true solution types, colloidal types, particulate dosage forms, suspension dosage forms, etc. Semi-solid dosage forms can be ointments, creams, pastes, gels, etc. Solid dosage forms can be orally dissolving films, tablets, capsules, dripping pills, pills, powders, granules, suppositories, freeze-dried powder injections, inclusion compounds, implants, patches, etc.

[0041] The compounds of the present invention can be incorporated into the said dosage forms. This can be achieved in a known manner by mixing with pharmaceutically suitable carriers, excipients and / or other adjuvants.

[0042] The single administration dose of the drug of the present invention is 0.01 - 100 mg of active ingredient / kg body weight, preferably 0.02 - 80 mg of active ingredient / kg body weight, more preferably 0.05 - 50 mg of active ingredient / kg body weight, still more preferably 0.08 - 40 mg of active ingredient / kg body weight, particularly preferably 0.1 - 20 mg of active ingredient / kg body weight, also preferably 0.1 - 15 mg of active ingredient / kg body weight, for example 0.1 - 10 mg of active ingredient / kg body weight, 0.1 - 8 mg of active ingredient / kg body weight, and most preferably 0.2 - 6 mg of active ingredient / kg body weight.

[0043] In one embodiment, the drug of the present invention is administered at least once a month, for example, 1, 2, 3, 4 or 5 times a month. Preferably, the drug of the present invention is administered 1, 2 or 3 times a month. Here, the term "active ingredient" refers to the compound of formula (I) or its salt in the present invention.

[0044] plan

[0045] The first aspect of the present invention relates to a compound of formula (I) or its salt:

[0046]

[0047] Wherein:

[0048] R represents -OCH(R 1 )(R 2 ),

[0049] R 1 represents hydrogen or (C1-C8)-alkyl,

[0050] 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.

[0051] In a preferred embodiment, R1 represents hydrogen and R2 represents a 3- to 10-membered heterocyclic group, wherein the heterocyclic group contains one or more heteroatoms selected from O and N.

[0052] In a preferred embodiment, R 1 represents hydrogen or (C1-C6)-alkyl, and R 2 represents a 3- to 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, and R 2 represents a 3- to 8-membered heterocyclic group, wherein the heterocyclic group contains 1 to 3 heteroatoms selected from O and N.

[0053] In a preferred embodiment, the 3- to 8-membered heterocyclic group is a 3- to 8-membered heterocyclic group containing 1 to 3 (preferably 1 to 2) O atoms, 1 to 3 (preferably 1 to 2) N atoms and / or 1 to 3 (preferably 1 to 2) S atoms.

[0054] In a further preferred embodiment, R 1 represents hydrogen, and R 2 represents a 5- to 7-membered heterocyclic group, wherein the heterocyclic group contains 1 to 3 heteroatoms selected from O and N.

[0055] In a further preferred embodiment, R 1 represents hydrogen, and R 2 represents morpholinyl, preferably morpholin-3-yl.

[0056] The definitions of the groups listed above in general terms or within the preferred ranges can be combined with each other as needed, i.e., including combinations between the given preferred ranges.

[0057] Highly particularly preferred compounds of the present invention are .

[0058] The second aspect of the present invention provides a method for preparing a compound of formula (I) or a salt thereof, the method comprising:

[0059]

[0060] In the presence of a catalyst, a condensing agent and a condensation activator, porphyrinogen (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 compounds of general formula (II)

[0061] RH (II)

[0062] wherein R is as defined above.

[0063] The catalyst applicable to 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-dimethylpyridine or a mixture thereof, more preferably DIPEA, NMI, and particularly preferably DIPEA.

[0064] The condensing agent applicable to 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, and more preferably EDCI.

[0065] Examples of the condensation activator applicable to the method of the present invention include but are not limited to N,N-dimethylaminopyridine (DMAP), 4-pyrrolidinopyridine (4-PPY), 1-hydroxy-7-azabenzotriazole (HOAT), 1-hydroxybenzotriazole (HOBT), N-hydroxysuccinimide (NHS), N-hydroxyphthalimide (NHPI), pentafluorophenol (PFP), etc., and preferably HOBT.

[0066] The polar organic solvent applicable to the method of the present invention can be preferably selected from N,N-dimethylformamide (DMF), N,N-dimethylacetamide, formamide, dimethyl sulfoxide (DMSO), acetone, pyridine or a mixture thereof, and more preferably DMF.

[0067] 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).

[0068] In the method of the present invention, the molar ratio of the compound having a reactive hydrogen atom to the condensing agent or the condensation activator is 1:(0.6 - 1.5), preferably 1:(0.7 - 1.3), and more preferably 1:(0.8 - 1.1).

[0069] In a preferred embodiment of the present invention, the compound having a reactive hydrogen atom is tert-butyl (R)-3-(hydroxymethyl)morpholine-4-carboxylate.

[0070] 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-butoxycarbonyl (Boc). The removal of the amino protecting group can be carried out by conventional methods known to those skilled in the art. For example, it can be removed using a solution of hydrogen chloride in a specific organic solvent (such as dioxane, ethyl acetate, methanol), preferably a hydrogen chloride dioxane solution.

[0071] The method of the present invention for preparing the compound of formula (I) uses a combination of a specific organic base catalyst, a condensing agent, and a condensation activator, particularly a combination of DIPEA, EDCI, and HOBT in a specific ratio. The method can obtain the target compound of high purity in good yield through simple post-treatment. Excessively low or high amounts of the catalyst, condensing agent, or condensation activator can adversely affect the target product, such as making post-treatment difficult, increasing the proportion of by-products, and reducing the yield of the target product.

[0072] 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.

[0073] In the above-described method for preparing the compounds of the present invention, the method may optionally include additional post-processing steps. These post-processing steps may include, for example, pH adjustment, crystallization, extraction, filtration, concentration under reduced pressure, and drying, among other conventional purification steps. Each of these steps can be performed in conventional manners known to those skilled in the art. If present, extraction is typically performed using a mixture of dichloromethane and methanol, preferably a mixture of dichloromethane and methanol at a ratio of 10:1 (v / v). Drying is typically performed by freeze drying, infrared drying, vacuum drying, or the like, with freeze drying being preferred.

[0074] In a preferred embodiment of the present invention, the method may further comprise a purification step by chromatography. The purification may be performed using a normal phase silica gel column having a silica gel particle size of 30-100 μm, preferably 40-63 μm, a loading of 20-120 g, preferably 40 g, and elution using dichloromethane / methanol containing 5% acetic acid (v / v) (elution gradient 100% / 0% to 90% / 10%, gradient elution time 15 minutes); or a reverse phase cyano column having a particle size of 10-100 μm, preferably 20-45 μm, a loading of 20-120 g, preferably 40 g, and elution using water containing 5% acetic acid / acetonitrile containing 5% acetic acid (v / v) (elution gradient 95% / 5% to 5% / 95%, gradient elution time 20 minutes).

[0075] 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.

[0076] 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.

[0077] The fourth aspect of the present invention relates to the 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.

[0078] The present invention also relates to a method for inhibiting Gram-negative or Gram-positive cocci, bacilli or coccobacilli, which method comprises contacting the compound or a salt thereof with Gram-negative or Gram-positive cocci, bacilli or coccobacilli and irradiating with light of a specific wavelength in an inhibitory effective amount. Preferably, the inhibition is for non-therapeutic purposes. The contacting can be carried out in vivo or in vitro, for example, in vitro.

[0079] 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 method comprises administering to a subject in need a therapeutically effective amount of the compound or a salt thereof and irradiating with light of a specific wavelength in a therapeutically effective amount. Preferably, the subject is a mammal, more preferably a human.

[0080] The compound or a salt thereof according to the first aspect of the present invention, as a photosensitizer for use in photodynamic therapy, specifically for treating a disease or condition caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli.

[0081] The Gram-negative or Gram-positive cocci, bacilli or coccobacilli may 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.

[0082] In a preferred embodiment, the Gram-negative or Gram-positive cocci, bacilli or coccobacilli are Porphyromonas gingivalis ( Porphyromonas gingivalis ), Actinobacillus actinomycetemcomitans ( Actinobacillus actinomycetemcomitans ), Tannerella forsythia ( Tannerella forsythia ), Fusobacterium nucleatum ( Fusobacterium nucleatum ), Prevotella intermedia ( Prevotella intermedia ), Pseudomonas aeruginosa ( Pseudomonas aeruginosa ), or Staphylococcus aureus ( Staphylococcus aureus ); preferably Porphyromonas gingivalis.

[0083] In a preferred embodiment, the disease or condition is selected from one or more of the following: periodontitis, gingivitis, dental plaque and dental caries; preferably, the periodontitis is chronic periodontitis or aggressive periodontitis;

[0084] Preferably, the disease or condition is selected from one or more of the following: gingival swelling, gingival bleeding, gingival pain, halitosis, periodontal pocket formation, alveolar bone resorption and tooth loosening.

[0085] 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.

[0086] 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.

[0087] 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.

[0088] 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.

[0089] Preferably, the optical power of the light irradiation is 30 mW-1000 mW, preferably 60 mW-600 mW, more preferably 80 mW-300 mW.

[0090] 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.

[0091] 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.

[0092] Preferably, in the context of the present invention, the salt is a pharmaceutically acceptable salt.

[0093] A 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, optionally further comprising one or more other active compounds. The other active compounds are compounds that are generally also effective in treating diseases or conditions caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli.

[0094] In one embodiment, the pharmaceutical composition is a kit, which further comprises instructions for using the compound or its salt as a photosensitizer in photodynamic therapy.

[0095] 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, still further preferably 4 mg - 100 mg of the compound of formula (I) or its salt, such as 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.

[0096] In a preferred embodiment, the drug further comprises a pharmaceutically acceptable carrier, excipient and / or other adjuvants. When a pharmaceutically acceptable carrier, excipient and / or other adjuvants are included, 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 usually combined to form a suitable administration form or dosage form, and this procedure includes mixing, granulating, compressing, dissolving or freeze-drying the components by suitable methods. The content of the carrier in the drug can be 1 to 98% by weight, usually about 80% by weight. For convenience, other adjuvants such as local anesthetics, preservatives, buffers, etc. can be directly dissolved in the carrier.

[0097] Example

[0098] Detailed synthetic examples of the selected compounds of the present invention are given below. However, these examples are only exemplary and should not be construed as limiting the scope of the present invention in any way.

[0099] Recorded in the form of an NMR peak list in the synthetic examples 1 The 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 solvent used in each case is also specified in the table.

[0100] In the present invention, in addition to the above recorded in the form of an NMR peak list 1In addition to the \(^1H\) NMR spectral data, the structures of the compounds prepared in the synthesis examples were also characterized by liquid chromatography-mass spectrometry (LC-MS). In the present invention, the liquid chromatography-mass spectrometer (LC-MS) used for detecting mass spectrometry data was purchased from Waters Technology (Shanghai) Co., Ltd., model: SQD2; the medium-pressure column chromatography machine used for the separation and purification of compounds was purchased from Santai Technology (Changzhou) Co., Ltd., model: SepaBean mechine T.

[0101] Unless otherwise specified or defined, the scientific and technical terms used in conjunction with the present invention will have the meanings commonly understood by those of ordinary skill in the art.

[0102] For the reagents or instruments whose manufacturers are not indicated, they are all commercially available conventional products commonly used in the art. In the present invention, unless otherwise stated, all operations are carried out at room temperature and atmospheric pressure. Unless otherwise specified, the contents and percentages in the context of this application are based on weight.

[0103] A. Synthesis Examples

[0104] Example 1: Synthesis of D-01

[0105] Protoporphyrin (200 mg, 0.36 mmol, 1 equ) was dissolved in 10 mL of 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. After stirring at room temperature for 10 minutes, tert-butyl (R)-3-(hydroxymethyl)morpholine-4-carboxylate (78 mg, 0.36 mmol, 1.0 equ) was added, and the mixture was continuously stirred at room temperature for 4 hours. Then 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 a normal-phase silica gel column (dichloromethane / methanol containing 5% acetic acid (v / v) 100% / 0% to 90% / 10%, elution time 15 min). After evaporating the solvent, it was lyophilized to obtain 149 mg of intermediate D-01-01, with a yield of 55%.

[0106] 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 detecting the completion of the reaction by liquid chromatography-mass spectrometry, it was concentrated under reduced pressure at room temperature and lyophilized to obtain product D-01 (124 mg, yield 95%).

[0107] LC-MS (m / z): 662.6 [M+H] + 。

[0108] 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).

[0109] B. Absorption spectra of the compounds of the present invention

[0110] To explore the optimal excitation wavelength of the drug, the absorption wavelength of the compound of the present invention was detected. The specific steps are as follows:

[0111] Prepare a dimethyl sulfoxide (brand: Shanghai Runjie) solution of the porphyrin derivative D-01 of the present invention with a concentration of 20 μg / mL, 4 mL each, and place it in a cuvette. Use an ultraviolet spectrophotometer (purchased from Shanghai Yuanxi Instrument Co., Ltd., model: X-8S) to test the absorption spectrum of the solution in the cuvette.

[0112] As Figure 1 shown, the compound of the present invention can absorb the energy at 405, 505, 540, 575 and 630 nm, and especially absorbs the strongest energy near 405 nm. Based on the above results, the excitation wavelengths of the porphyrin derivative of the present invention were determined to be 405 nm and 630 nm. Among them, the peak shape of protoporphyrin is not smooth, indicating that there is molecular aggregation at this concentration. At the same time, it indicates 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.

[0113] C. Effect Example

[0114] Example 1

[0115] 1. Materials and methods

[0116] 1.1 Information of the drug to be tested

[0117] Test drug: Compound D-01 of the present invention prepared in Synthesis Example 1.

[0118] Positive drug 1: Protoporphyrin (PPIX).

[0119] Positive drug 2: 5-aminolevulinic acid (5-ALA).

[0120] Solvent: DMSO.

[0121] 1.2 Experimental methods

[0122] After resuscitating Porphyromonas gingivalis (purchased from Beijing Beina Chuanglian Biotechnology Research Institute, batch number BNCC353909), inoculate it into BHI medium (brain heart infusion medium) and culture it to the logarithmic phase under anaerobic conditions of 10% H2, 10% CO2, 80% N2, and 37°C; take 1 mL of the bacterial solution, measure the OD value, centrifuge and discard the supernatant, and then add a certain amount of BHI medium to adjust the bacterial solution concentration to 10 10 CFU / mL (1 OD≈9.52*10 9 CFU / mL) for standby.

[0123] Test drug group: Add 100 μL of the bacterial solution, 898 μL of BHI medium, and 2 μL of the DMSO solution of the test drug to the centrifuge tube respectively, and shake well (to make the final concentration of the bacterial solution 10 9 CFU / mL, the final concentration of the test drug is 10 μg / mL, the final concentration of positive drug 1 is 10 μg / mL, and the final concentration of positive drug 2 is 80 μg / mL). Protect from light and culture for 4 h under the same anaerobic conditions as above, then centrifuge and discard the supernatant, and then resuspend with 1 mL of BHI medium. Add the resuspended solution to a 96-well plate, add 100 μL of the bacterial solution to each well, and repeat 3 times for each group, with 4 replicates each time. Irradiate with a 405 nm laser, the laser output power is 300 mW, irradiate for 66.67 s, and the light dose is 20 J. After the irradiation is completed, continue to culture for 24 h under the same anaerobic conditions as above, and then measure the OD value of each group of bacterial solutions at 600 nm and calculate the antibacterial rate.

[0124] Blank light group (abbreviated as blank light group): Carry out according to the above method, the difference is: add 100 μL of the bacterial solution and 900 μL of BHI medium to the centrifuge tube.

[0125] Blank non-irradiation group (abbreviated as blank non-group): Carry out according to the above method, the difference is: add 100 μL of the bacterial solution and 900 μL of BHI medium to the centrifuge tube, without irradiation treatment.

[0126] Solvent group: Carry out according to the above method, the difference is: add 100 μL of the bacterial solution, 898 μL of BHI medium, and 2 μL of DMSO to the centrifuge tube.

[0127] The formula for calculating the antibacterial rate is as follows:

[0128]

[0129] Among them: OD 阴性 represents the average value of the OD value of the bacterial liquid measured in the blank light irradiation group; OD 待测 represents the average value of the OD value of the bacterial liquid measured in the test drug group.

[0130] 1.3 Calculation of statistical differences

[0131] Use GraphPad Prism 8 for statistical analysis and compare the OD 600 value differences to evaluate the antibacterial effect. One-way analysis of variance (One-way ANOVA) is used for multi-group comparison analysis, and post hoc multiple comparisons are performed. P < 0.05 is considered to have statistical differences. Among them, * < 0.05, ** < 0.01, *** ＜ 0.001, **** < 0.0001, representing a significant decrease.

[0132] 2. Results and discussion

[0133] 2.1 Results of the first biological replication

[0134] The results are as shown in Figure 2 (a). Compared with the blank light irradiation group, the optical density of compound D-01 and the optical density of positive drug 1 were significantly decreased (p < 0.0001), and their antibacterial rates were 64.14% and 32.65% respectively.

[0135] 2.2 Results of the second biological replication

[0136] The results are as shown in Figure 2 (b). Compared with the blank light irradiation group, the optical density of compound D-01 and the optical density of positive drug 1 were significantly decreased (p < 0.0001). The antibacterial rates of D-01 and positive drug 1 were 72.5% and 28.2% respectively.

[0137] 2.3 Results of the third biological replication

[0138] The results are as shown in Figure 2 (c). Compared with the blank light irradiation group, the optical density of compound D-01 and the optical density of positive drug 1 were significantly decreased (p < 0.0001). The antibacterial rates of D-01 and positive drug 1 were 68.2% and 32.4% respectively.

[0139] 2.4 Antibacterial rates of three biological replication experiments

[0140] The results are as shown in Figure 2 (d) and Table 1. The compound D-01 of the present invention has a significant and relatively stable antibacterial effect, and the effects are significantly better than those of the control compounds PPIX and 5-ALA, with significant differences (p < 0.0001).

[0141] Table 1: Inhibitory effects of compounds D-01, PPIX, and 5-ALA on Porphyromonas gingivalis

[0142] .

Claims

1. A compound of formula (I) or a salt thereof: (I) Wherein: R represents -OCH(R 1 )(R 2 ), R 1 represents hydrogen, R 2 represents a saturated monocyclic heterocyclic group having 2, 3, 4, 5 or 6 carbon atoms and 1 or 2 heteroatoms selected from oxygen and nitrogen.

2. The compound or a salt thereof according to claim 1, wherein: R 1 represents hydrogen, R 2 represents a saturated monocyclic heterocyclic group having 2, 3, 4, 5 or 6 carbon atoms and 2 heteroatoms selected from oxygen and nitrogen.

3. The compound or its salt according to claim 2, wherein the compound is .

4. A method for preparing the compound of formula (I) or a salt thereof according to any one of claims 1 to 3, the method 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 condensing agent, and a condensation activator, wherein the compound having a reactive hydrogen atom is selected from compounds 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-dimethylpyridine, 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-pyrrolidinopyridine, 1-hydroxy-7-azabenzotriazole, 1-hydroxybenzotriazole, N-hydroxysuccinimide, N-hydroxyphthalimide, pentafluorophenol; or The molar ratio of the compound having a reactive hydrogen atom to the condensing agent or the condensation activator is 1:(0.6 - 1.5).

6. Use of the 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 the compound of formula (I) or a salt thereof according to any one of claims 1 to 3 in the preparation of a medicament for the treatment of a disease or condition caused by Porphyromonas gingivalis.

8. 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 dental caries; or the disease or condition is selected from one or more of the following: gingival swelling, gingival bleeding, gingival pain, bad breath, periodontal pocket formation, alveolar bone resorption, and tooth loosening.

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

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