Mono-substituted amide porphyrin derivative, preparation method thereof and application of mono-substituted amide 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 an efficient photosensitizer for treating hyperproliferative diseases such as cancers or precancerous lesions in photodynamic therapy, or treating oral diseases # imgabs0 # (I) such as periodontitis, gingivitis, dental plaque and decayed teeth caused by bacteria such as Gram-negative anaerobic bacillus such as porphyromonas gingivalis.

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 amide 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] Cervical cancer is one of the most common tumors of the female reproductive system and the second leading cause of cancer-related death among women worldwide. Currently, only 5-aminolevulinic acid and 5-aminolevulinic acid hexyl ester (HAL) are used as photosensitizers for photodynamic therapy of cervical precancerous lesions in the world, and they are in the clinical trial stage. There are no photosensitizer drugs for the treatment of cervical cancer or precancerous lesions on the market, so it is urgent to develop such new photosensitizers to promote the development of the industry.

[0005] 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.

[0006] In summary, it is of great significance to develop photosensitizers with high efficiency in killing cancer cells and / or antibacterial ability. Summary of the invention

[0007] The present invention aims to provide a class of monosubstituted amide porphyrin derivatives, a preparation method thereof and use thereof as photosensitizers, and more specifically to use them in photodynamic therapy to treat hyperproliferative diseases, especially cervical cancer or cervical precancerous lesions, or to inhibit bacteria to treat diseases such as periodontitis, gingivitis, dental plaque or caries.

[0008] The first aspect of the present invention relates to a compound of formula (I) or a salt thereof: (I) in: R stands for -NR 1 R 2 , R 1 and R 2 Each independently represents hydrogen, heterocyclylalkyl or heteroarylalkyl, wherein R 1 and R 2 Different from hydrogen, wherein the heterocyclic or heteroaryl group comprises one or more heteroatoms selected from O, S and N; The condition is to exclude compounds and .

[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 the compound or salt thereof according to the first aspect of the present invention in the preparation of a medicament for treating a hyperproliferative disease.

[0012] The present invention also relates to a method for treating a hyperproliferative disease, which comprises administering a therapeutically effective amount of a compound of formula (I) or a salt thereof as described above to a subject in need thereof, and administering a therapeutically effective amount of light irradiation of a specific wavelength, preferably, wherein the subject is a mammal, more preferably a human.

[0013] The fifth 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.

[0014] 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.

[0015] 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.

[0016] The sixth aspect of the present invention relates to a pharmaceutical composition, which comprises the compound or salt thereof according to the first aspect of the present invention, and optionally further comprises one or more other active compounds.

[0017] Surprisingly, the present invention has the following beneficial effects: The present invention develops a novel class of monosubstituted amide 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 to treat hyperproliferative diseases such as cancer or precancerous lesions, especially cervical cancer or cervical precancerous lesions, and especially when the dosage is significantly smaller, its efficacy is significantly better than that of the drug 5-ALA, and is equivalent to or even significantly better than that of the drug HAL, and 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

[0018] Figure 1 The standard curve of the transdermal diffusion amount of PPIX and the compound of the present invention in the in vitro transdermal permeation experiment is shown.

[0019] Figure 2The standard curve of tissue retention samples of PPIX and the compounds of the present invention for 6 hours in the in vitro percutaneous permeation experiment is shown.

[0020] Figure 3 The 6-hour unit area tissue retention of PPIX and the compound of the present invention is shown. Compared with the PPIX group, the compound N-01 group *P<0.05, the compound N-02 group ****P<0.0001.

[0021] Figure 4 The weight changes of mice in each treatment group after administration are shown. The data points represent the mean weight within the group, and the error bars represent the standard error (SEM).

[0022] Figure 5 The in vivo imaging images of mice in the high-dose groups of compounds N-01 and N-02 of the present invention are shown.

[0023] Figure 6 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).

[0024] Figure 7 It is the ultraviolet absorption spectrum of compounds N-01 and N-02 of the present invention. DETAILED DESCRIPTION

[0025] 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.

[0026] 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.

[0027] The term "optionally" as used herein 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.

[0028] Unless otherwise defined, the names of chemical groups are generally to be understood in such a way that the connection to the skeleton or the rest of the molecule is via the structural element of the last mentioned chemical group concerned, i.e., for example in the case of heterocyclyl-(C1-C8)-alkyl, via a carbon atom of the alkyl group. 1 R 2 In the case of 1 R 2 In the case of , the bond is through a nitrogen atom.

[0029] 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.

[0030] Unless otherwise defined, the term "heteroaryl" refers to a monocyclic, bicyclic or tricyclic heterocyclic group of carbon atoms and at least one heteroatom selected from oxygen, nitrogen and sulfur, wherein at least one ring is aromatic. Preferably, the heteroaryl is a 5-12 membered heteroaryl, for example containing 3, 4, 5, 6, 7 or 8 carbon atoms, which can be attached to the parent molecular moiety through any carbon atom or nitrogen atom contained in the heterocyclic ring. Examples include, but are not limited to, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-triazolyl, 1,2,5-triazolyl, 1,2,3-oxadi ...3-oxadiazolyl, 1,2,4-triazolyl, 1,3,4-triaz oxadiazole, 1,2,4-triazine, 1,3,5-triazine, tetrazolyl, benzofuranyl, benzisofuranyl, benzothiophenyl, benzisothiophenyl, indolyl, isoindolyl, indazolyl, benzothiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, 2,1,3-benzoxadiazole, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, benzotriazinyl, purinyl, pteridinyl, imidazopyridinyl, thienopyrimidinyl, thienopiperidinyl, and the like.

[0031] 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.

[0032] Unless otherwise defined, the term "heteroarylalkyl" is understood to mean a combination of the groups "heteroaryl" and "alkyl" as defined according to the invention, wherein the group is usually connected to the backbone or the remainder via the alkyl group, such as 5-12 membered heteroaryl-(C1-C8)-alkyl, 5-10 membered heteroaryl-(C1-C6)-alkyl and 5-6 membered heteroaryl-(C1-C4)-alkyl. Examples include, but are not limited to, pyrrolylmethyl, pyrrolylethyl, benzothienylmethyl, benzothienylethyl, furanylmethyl, furanylethyl, benzofuranylmethyl, thienylmethyl, thienylethyl, pyridylmethyl, etc.

[0033] Unless otherwise defined, the term "heterocyclylalkyl" is understood to mean a combination of the groups "heterocyclyl" and "alkyl" defined according to the invention, wherein the group is usually connected to the backbone or the remainder via the alkyl group, such as 3-10 membered heterocyclyl-(C1-C8)-alkyl, 3-8 membered heterocyclyl-(C1-C6)-alkyl and 5-6 membered heterocyclyl-(C1-C4)-alkyl. Examples include, but are not limited to, 2-(morpholin-4-yl)ethyl, morpholin-3-ylmethyl, etc.

[0034] 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.

[0035] As used herein, the term "cancer" includes, but is not limited to, breast cancer, respiratory tract cancer, brain cancer, reproductive organ cancer, digestive tract cancer, urinary tract cancer, liver cancer, eye cancer, skin cancer, head and neck tumors and distant metastases, and also includes multiple myeloma, lymphoma and sarcoma.

[0036] Unless otherwise clearly defined, "precancerous lesions" in this article refer to abnormal cell proliferation with a high possibility of becoming cancerous. Examples include but are not limited to cervical precancerous lesions, oral leukoplakia, myelodysplastic diseases, familial intestinal polyps, skin nevi, psoriasis, solar keratosis, etc.

[0037] As used herein, the term "treating" means killing, inhibiting or slowing the growth or increase in size of a hyperproliferative cell mass or group or a tumor or cancerous growth, reducing the number of hyperproliferative cells, or preventing spread to other anatomical sites, as well as methods of reducing the size of a hyperproliferative growth or the number of hyperproliferative cells. However, it should be understood that "treating" does not necessarily mean curing or completely eliminating a hyperproliferative growth.

[0038] 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.

[0039] 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.

[0040] Preferred are oral mucosal administration, oral administration, intramuscular injection, topical administration, vaginal administration, rectal administration, intraperitoneal administration or intravenous administration. Oral mucosal administration includes local injection, local coating, local washing, oral gargle, local controlled / slow release, microneedle and other device administration.

[0041] 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.

[0042] For these administration routes, the compounds according to the invention can be administered in suitable administration forms.

[0043] 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.

[0044] 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.

[0045] 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.

[0046] 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.

[0047] plan The first aspect of the present invention relates to a compound of formula (I) or a salt thereof: (I) in: R stands for -NR 1 R 2 , R 1 and R 2 Each independently represents hydrogen, heterocyclylalkyl or heteroarylalkyl, wherein R 1 and R 2 Not at the same time hydrogen; wherein the heterocyclic group or heteroaryl group comprises one or more heteroatoms selected from O, S and N; preferably wherein the heterocyclic group or heteroaryl group comprises one or two heteroatoms selected from O, S and N; The condition is to exclude compounds and .

[0048] In a preferred embodiment, R 1 and R 2 Each independently represents hydrogen, 3-10 membered heterocyclyl(C1-C8)-alkyl or 5-12 membered heteroaryl(C1-C8)-alkyl.

[0049] In a further preferred embodiment, R 1 and R 2Each independently represents hydrogen, 3-8 membered heterocyclyl(C1-C6)-alkyl or 5-10 membered heteroaryl(C1-C6)-alkyl.

[0050] In a preferred embodiment, the 3-8 membered heterocyclyl (C1-C6)-alkyl and the 5-10 membered heteroaryl (C1-C6)-alkyl are 3-8 membered heterocyclyl (C1-C2)-alkyl and 5-10 membered heteroaryl (C1-C2)-alkyl containing 1-3 (preferably 1-2) O atoms, 1-3 (preferably 1-2) N atoms and / or 1-3 (preferably 1-2) S atoms, respectively.

[0051] In a further preferred embodiment, R 1 and R 2 Each independently represents hydrogen, morpholino(C1-C6)-alkyl or thiophene(C1-C6)-alkyl.

[0052] In a further preferred embodiment, R 1 and R 2 Each independently represents hydrogen, morpholin-4-yl(C1-C6)-alkyl (preferably 2-(morpholin-4-yl)ethyl), morpholin-3-yl(C1-C6)-alkyl (preferably morpholin-3-ylmethyl) or thiophen-2-yl(C1-C6)-alkyl (preferably thiophen-2-ylmethyl).

[0053] 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.

[0054] Very particular preference is given to the compounds of the formula (I) according to the invention listed in Table 1 below.

[0055] Table 1: Compounds of formula (I) in which R has the meaning given below.

[0056] .

[0057] Note:" " indicates connection with the rest of the compound of formula (I).

[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] 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.

[0060] 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.

[0061] 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.

[0062] 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.

[0063] 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.

[0064] 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).

[0065] 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).

[0066] In a preferred embodiment of the present invention, the compound having a reactive hydrogen atom is selected from tert-butyl 3-(aminomethyl)morpholine-4-carbonate or 2-morpholino-N-(thiophen-2-ylmethyl)ethylamine.

[0067] 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.

[0068] 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.

[0069] 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.

[0070] 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.

[0071] 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).

[0072] 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.

[0073] 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.

[0074] The fourth aspect of the present invention relates to use of the compound or salt thereof according to the first aspect of the present invention in the preparation of a medicament for treating a hyperproliferative disease.

[0075] The present invention also relates to a method for treating a hyperproliferative disease, which comprises administering a therapeutically effective amount of a compound of formula (I) or a salt thereof as described above to a subject in need thereof, and administering a therapeutically effective amount of light irradiation of a specific wavelength, preferably, wherein the subject is a mammal, more preferably a human.

[0076] 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 hyperproliferative diseases.

[0077] Preferably, the hyperproliferative disease is cancer or a precancerous lesion, and the cancer is preferably selected from bladder cancer, esophageal cancer, bronchial cancer, oral cancer, nasopharyngeal cancer, liver cancer, pancreatic cancer, skin cancer, penile cancer, cervical cancer, vaginal cancer, endometrial cancer, ovarian cancer, colorectal cancer, kidney cancer, urothelial cell cancer, thyroid cancer, breast cancer, anal cancer, Kaposi's sarcoma, lung cancer, gastric cancer, bile duct cancer, prostate cancer, melanoma and brain cancer, more preferably cervical cancer and skin cancer, particularly preferably cervical cancer; the precancerous lesion is preferably selected from cervical precancerous lesions, oral leukoplakia, myelodysplastic disease, familial intestinal polyps, skin nevus, psoriasis, solar keratosis, more preferably cervical precancerous lesions and skin nevus, particularly preferably cervical precancerous lesions.

[0078] The fifth 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.

[0079] 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.

[0080] 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.

[0081] 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.

[0082] Preferably, 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.

[0083] 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.

[0084] 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.

[0085] 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.

[0086] In a preferred embodiment, the compound of formula (I) or a salt thereof is used as a photosensitizer in photodynamic therapy to treat cancer or precancerous lesions, in particular cervical cancer or cervical precancerous lesions, or for treating diseases or conditions caused by Gram-negative or positive cocci, bacilli or coccobacilli.

[0087] 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.

[0088] 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.

[0089] 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; when used to treat hyperproliferative diseases, 630 nm is preferred; when used to treat diseases or conditions caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli, 405 nm is preferred.

[0090] Preferably, the optical power of the light irradiation is 30 mW-1000 mW, preferably 60 mW-600 mW, more preferably 80 mW-300 mW.

[0091] 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.

[0092] 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.

[0093] Preferably, in the context of the present invention, the salt is a pharmaceutically acceptable salt.

[0094] The sixth 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. The other active compounds are compounds that are generally effective in treating hyperproliferative diseases, or diseases or conditions caused by Gram-negative or Gram-positive cocci, bacilli or coccobacilli.

[0095] 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.

[0096] 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.

[0097] 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.

[0098] 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.

[0099] The NMR peaks reported in the synthetic examples are 1 H NMR spectral data were obtained on a Bruker 600 MHz NMR spectrometer and the signals listed have the meanings given below: s = singlet, d = doublet, m = multiplet, brs = broad singlet. The deuterated solvents used in each case are also specified in the table.

[0100] In the present invention, in addition to the above-mentioned NMR peak list form 1In 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.

[0101] 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.

[0102] 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.

[0103] A. Synthesis Examples Example 1: Synthesis of N-01 Protoporphyrin (200 mg, 0.36 mmol, 1 equivalent (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. After stirring for 10 minutes, tert-butyl 3-(aminomethyl)morpholine-4-carbonate (78 mg, 0.36 mmol, 1 equ) was added and continued to stir at room temperature for 5 hours. The reaction solution was poured into 30 mL of water and extracted with 3× 30 mL of a mixed solution of dichloromethane / methanol (10 / 1, v / v). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The product was purified by normal phase silica gel column (dichloromethane / methanol containing 5% acetic acid (v / v) 100% / 0% to 90% / 10%, elution time 15 min). After evaporation of the solvent, the intermediate N-01-01 (123 mg, yield 45%) was obtained by lyophilization.

[0104] 123 mg of intermediate N-01-01 was dissolved in 5 mL of 4 mol / L hydrogen chloride dioxane solution, stirred for 2 hours, concentrated under reduced pressure, and lyophilized to obtain the target compound N-01 (101 mg, yield 95%).

[0105] LC-MS: 661.7 [M+H] + .

[0106] 1H NMR (600 MHz, CDCl3): δ 11.02 (s, 1H), 10.69 (s, 1H), 10.64-10.63(m, 2H), 8.20-8.13 (m, 2H), 6.48-6.44 (m, 4H), 4.47-4.40 (m, 4H), 3.74-3.70(m, 6H), 3.67-3.62 (m, 6H), 3.61-3.60 (m, 1H), 3.28-3.10 (m, 10H), 2.69-2.62(m, 2H).

[0107] Example 2: Synthesis of N-02 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. After stirring for 10 minutes, 2-morpholino-N-(thiophen-2-ylmethyl)ethylamine (81 mg, 0.36 mmol, 1.0 equ) was added, and stirring was continued at room temperature for 2 hours. The product was purified by reverse phase cyano column (water containing 5% acetic acid / acetonitrile containing 5% acetic acid (v / v) 95% / 5% to 5% / 95%, elution time 20 min), concentrated under reduced pressure, and lyophilized to obtain the target compound N-02 (200 mg, yield 73%).

[0108] LC-MS: 771.8 [M+H] + .

[0109] 1 H NMR (600 MHz, DMSO- d 6): δ 10.08-9.97 (m, 4H), 8.42-8.35 (m, 2H), 7.32-7.28 (m, 0.5H), 7.20 (d, J= 4.8 Hz, 0.5H), 6.90 (s, 0.5H), 6.86 (s, 1H), 6.67 (br s, 0.5H), 6.41-6.37 (m, 2H), 6.21-6.17 (m, 2H), 4.68-4.65 (m, 2H),4.34-4.28 (m, 4H), 3.64-3.61 (m, 6H), 3.56 (s, 3H), 3.52 -3.51 (m, 3H), 3.29-3.26 (m, 3H), 3.19-3.17 (m, 3H), 3.10-3.05 (m, 10H),-4.39 (s, 2H).

[0110] B. Absorption spectrum of the compound 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 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.

[0111] like Figure 7 As shown, the absorption wavelengths of the compounds of the present invention are similar, that is, they 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 wavelengths of the series of porphyrin derivatives of the present invention are determined to be 405 nm and 630 nm. Among them, the peak shape of protoporphyrin is not smooth, indicating that the molecules have molecular aggregation at this concentration. At the same time, it is suggested that the ACQ effect of the compounds 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.

[0112] C. Effect Example C.1 Treatment of hyperproliferative diseases Information of the drug to be tested: Compounds N-01 and N-02 of formula (I) of the present invention are shown in Table 1 above.

[0113] Compound: Protoporphyrin (PPIX).

[0114] Positive drug 1: 5-aminolevulinic acid (5-ALA).

[0115] Positive drug 2: 5-aminolevulinic acid hexyl ester (HAL) hydrochloride.

[0116] The glue matrix used in the present invention is prepared by the following method: Add 90 mL of pure water to a 200 mL beaker and place it on a magnetic stirrer. Slowly add 1.5 g of sodium carboxymethyl cellulose under stirring. After stirring at room temperature for 40 minutes, heat to 40 ° C, continue stirring for 30 minutes, and then cool to room temperature under stirring. Then, add 0.5 g of laurocapram, and adjust the pH to about 10 with an appropriate amount (about 2 mL) of 1 mol / L sodium hydroxide solution. Add an appropriate amount of pure water so that the total weight of the contents in the beaker is 100 g, stir evenly, and leave overnight to obtain a transparent blank slurry matrix.

[0117] Example 1: Transdermal penetration experiment In order to evaluate the effectiveness of the compounds of the present invention and provide a basis for the in vivo experimental administration method, the in vitro transdermal absorption and intradermal retention of the compounds of the present invention were tested. The specific steps are as follows: 1.1 Experimental methods A transdermal diffusion instrument with a single-chamber Franz cell was used, miniature pig skin was used as the in vitro transdermal absorption skin, and the concentration of the compound in the receiving solution at different time points was measured using an ELISA instrument, the cumulative permeation amount was calculated, and the amount of compound retained in the skin at the end of sampling was measured. The specific steps are as follows: (1) Preparation of test preparation: Accurately weigh 4 mg each of protoporphyrin, N-01 and N-02, dissolve them in 200 μL DMSO, add them to 4 g of the slurry matrix prepared above, vortex to mix, and prepare a test preparation with a concentration of 1 mg / g.

[0118] (2) Determination of fluorescence value of receiving pool sample The skin of miniature pigs was fixed in a Franz diffusion cell. A volume of V (about 15 mL) of PBS containing 40% PEG300 was added to the receiving cell. 3 g of the test preparation of 1 mg / g was added to the supply cell. The Franz diffusion cell was placed on a TP-6 ​​transdermal diffusion instrument (purchased from Tianjin Jingtuo Instrument Technology Co., Ltd.) and stirring (constant temperature 32°C, speed 150 rpm) was started and timing was started. 0.8 mL of samples were taken from the receiving cell at 0.5, 1, 2, 3, 4, and 6 hours (h) (immediately supplemented with 0.8 mL of PBS containing 40% PEG 300). The samples at all time points were placed in a 96-well plate, 100 μL per well, and 3 replicates were set for each sample. The fluorescence value Ft with 402 nm excitation and 631 nm reception was read on a microplate reader (BioTekSynergy H1).

[0119] (3) Construction of transdermal diffusion standard curve Protoporphyrin, N-01 and N-02 were dissolved in DMSO to prepare a 1 mg / mL stock solution. Protoporphyrin was diluted to 2, 1, 0.5, 0.25 and 0.125 ng / mL, and the compound of the present invention was diluted to 4, 2, 1, 0.5, 0.25 ng / mL using 40% PEG 300 in PBS. The fluorescence value F of each concentration was measured on an ELISA reader. s1 After regression analysis, record the standard curve equation y1 and determination coefficient R of transdermal diffusion 2 .

[0120] (4) Determination of fluorescence value of tissue retention samples After the experiment, the skin was cut into pieces, placed in a light-proof centrifuge tube, and 1 mL of DMSO was added. Ultrasound was performed at room temperature for 10 minutes. The samples were placed in a 96-well plate, with 100 μL per well and 3 replicates for each sample. The fluorescence value F was read on an ELISA reader at 402 nm excitation and 631 nm reception.

[0121] (5) Construction of standard curve for tissue retention samples Protoporphyrin, N-01 and N-02 were dissolved in DMSO to prepare 1 mg / mL stock solution. Protoporphyrin was diluted to 250, 125, 62.5, 31.25 and 15.625 ng / mL, and the compound of the present invention was diluted to 200, 100, 50, 25, 12.5, 6.25 ng / mL, and the fluorescence value F of each concentration was measured on an ELISA reader. s2 After regression analysis, record the standard curve equation y2 and determination coefficient R of tissue retention samples. 2 .

[0122] (6) Calculation of cumulative infiltration per unit area The fluorescence value Ft of the receiving pool sample at each time point is substituted into the transdermal diffusion standard curve equation y1 to calculate the transdermal diffusion concentration C at each time point. n , calculate the cumulative permeability per unit area Q (ng / cm) according to the following formula 1 2 ): (Formula 1) Among them, C n represents the drug concentration measured at the nth point (ng / mL), C i represents the drug concentration measured at the ith point (ng / mL), V represents the total volume of the receiving pool (mL), V0 represents the sampling volume per time (mL), and 1.13 represents the penetration area (cm 2 ).

[0123] (7) Calculation of tissue retention per unit area Substitute the fluorescence value F of the tissue retention sample into the tissue retention sample standard curve equation y2 to calculate the tissue retention sample concentration C. Calculate the tissue retention per unit area X (ng / cm) according to the following formula 2 2 ): (Formula 2) Where C represents the calculated drug concentration (ng / mL), V represents the DMSO extraction volume (mL), and 1.13 represents the permeation area (cm 2 ).

[0124] 1.2 Experimental Results Figure 1 The standard curves of transdermal diffusion of protoporphyrin and the compounds of the present invention are shown, R 2 The values ​​were all >0.98, indicating that the tested compounds had good linearity within the corresponding concentration range (protoporphyrin: 0-2 ng / mL; compounds of the present invention: 0-4 ng / mL).

[0125] Table 2 shows the cumulative permeation per unit area Q (ng / cm 2 ). The results show that the cumulative permeability per unit area of ​​N-01 is about 4 times that of PPIX, and the cumulative permeability per unit area of ​​N-02 is about 2.5 times that of PPIX.

[0126] Table 2: Cumulative permeation per unit area of ​​the tested compounds at different time points .

[0127] Figure 2 The tissue retention sample standard curves of protoporphyrin and the compounds of the present invention are shown, R 2 The values ​​were all >0.99, indicating that the tested compounds had good linearity within the corresponding concentration range (protoporphyrin: 15.625-250 ng / mL; compounds of the present invention: 6.25-200 ng / mL).

[0128] Figure 3 The intradermal tissue retention per unit area X (ng / cm2) of protoporphyrin and the compound of the present invention after 6 hours is shown. 2 ), indicating that the intradermal tissue retention per unit area of ​​the compounds of the present invention is significantly higher than that of protoporphyrin, among which N-02 has the largest tissue retention per unit area after 6 h, which is 130 ng / cm 2 (P<0.0001).

[0129] The transdermal permeation experiment showed that the compounds N-01 and N-02 of the present invention have better solubility and transdermal absorption than the lead compound PPIX, and can be used for in vivo efficacy tests by topical administration.

[0130] Example 2: Evaluation of drug efficacy in cervical carcinoma in situ in mice 2.1 Animal Experimental Methods (1) Cell culture Hela-luc human cervical cancer cells were cultured in vitro in RPMI-1640 medium with 10% fetal bovine serum in a 5% CO2 constant temperature incubator (Phcbi) at 37°C. Routine treatment and subculture were performed twice a week. When the cell saturation was 80%-90% and the number reached the required level, the cells were collected and counted (Countstar cell counter), and the cell density was adjusted to 5×10 with PBS. 7 / mL for later use, and then mix the Hela-luc cell suspension with the matrix gel in a 1:1 ratio.

[0131] (2) Animal vaccination 0.2 mL (5.0 × 10 6 Hela-luc cells (10 cells + matrix gel / mouse) were injected into the uterine cervix of Balb / c-nude mice (female, 6-8 weeks old, body weight 18-22 g). The model formation was observed on the 8th day after the injection of cancer cells using a mouse living imager (purchased from Shanghai Tianneng Technology Co., Ltd., model ABL-X5 PRO).

[0132] (3) Experimental groups and drug treatment At 2 weeks, 60 mice with appropriate tumor sizes were selected and randomly divided into 10 groups according to the tumor fluorescence intensity, with 6 mice in each group. The day of grouping was recorded as D0, and drug administration began on the day of grouping. Each group was administered once via vaginal application. The drug solution of each group was prepared according to Table 3 before the experiment and refrigerated in the dark before use.

[0133] Drug preparation and animal grouping were performed according to Table 3. Each group received topical administration via vaginal application, and 3 hours after administration, a 630nm semiconductor laser photodynamic therapy device (purchased from Guilin Xingda Optoelectronic Medical Device Co., Ltd., model PDT630-II) was used for treatment, with a light power of 100 mW, an irradiation time of 10 min, and a total energy of 60 J. During phototherapy, the optical fiber was inserted through the vagina to irradiate the cervical tumor site as completely as possible.

[0134] Table 3: Experimental group design and drug solution preparation table .

[0135] (4) Detection indicators After group administration, the tumor fluorescence intensity was measured by an imager twice a week, and the mice were weighed at the same time. The relative fluorescence intensity of the tumor was calculated according to the following formula 3: (Formula 3) Among them, Fi represents the average fluorescence intensity of the tumor in the experimental group on day i, F c It represents the average fluorescence intensity of tumor in blank control group on day i.

[0136] (5) Data processing and statistical analysis The number of days after grouping was used as the horizontal axis and the weight of the mice was used as the vertical axis. GraphPad Prism 8 was used to draw the graph. Statistical analysis was performed based on the data obtained at the end of the experiment to evaluate the differences between the groups. Ordinary one-way ANOVA was used in GraphPad Prism8 software for data analysis, and p<0.05 was considered to be significantly different.

[0137] 2.2 Experimental Results (1) Weight changes During the experiment, the weight changes of mice in each treatment group on days 0, 4, 7, 11 and 14 after administration were as follows: Figure 4 As shown, all treatment groups were well tolerated.

[0138] (2) Anti-tumor efficacy evaluation indicators The experiment was completed on the 14th day after grouping. Figure 5 The in vivo imaging of mice in the high-dose (4 mg / kg) treatment group of the compound of the present invention at different days after administration is shown. It can be clearly seen that at this dose, the tumor fluorescence intensity of N-01 and N-02 decreases with the increase in the number of days after treatment, indicating that the compound of the present invention has a significant inhibitory effect on tumor growth.

[0139] The tumor relative fluorescence intensity of each test compound calculated based on the tumor fluorescence intensity measured on days 4, 7, and 14 after grouping is summarized in Table 4.

[0140] Table 4: Relative fluorescence intensity of tumors in % for each group of tested compounds .

[0141] The above results show that on the 14th day after administration, the tumor growth trend of the illumination group alone was consistent with that of the blank control group, indicating that illumination alone had no significant inhibitory effect on the growth of Hela in situ tumors. Under the same illumination conditions, with the increase in the dosage of compounds N-01 and N-02 of the present invention (for example, at 1 mg / kg, 2 mg / kg and 4 mg / kg), the relative fluorescence intensity of the tumor decreased. Compared with the blank control group, the medium-dose (2 mg / kg) and high-dose (4 mg / kg) treatment groups of the compound of the present invention and the HAL (150 mg / kg) treatment group all had significant inhibitory effects on Hela in situ tumors, among which, the therapeutic effects of the compound of the present invention (medium-dose group 2 mg / kg) and HAL (150 mg / kg) were comparable, with no statistical difference (P>0.05); the therapeutic effect of the compound of the present invention (high-dose group 4 mg / kg) was significantly better than that of HAL (150 mg / kg), with statistical difference (P<0.05); in particular, the relative fluorescence intensity of the drug HAL (150 mg / kg) was 51.5% (P<0.05), in comparison, the relative fluorescence intensities of the compounds N-01 and N-02 (4 mg / kg) of the present invention were 21.0% (P<0.01) and 24.0% (P<0.01), respectively. Therefore, in terms of the overall tumor inhibition effect, the monosubstituted porphyrin derivatives of the present invention have an efficacy comparable to or even significantly superior to that of the drug HAL at a significantly smaller drug dosage.

[0142] In addition, during the experiment, the overall body weight of the mice was relatively stable and the animals were in good condition, indicating that the mono-substituted porphyrin derivatives of the present invention have good biosafety.

[0143] C.2 Antibacterial effect Example 3 1. Materials and Methods 1.1 Information on the drug to be tested Compound N-02 of formula (I) of the present invention shown in Table 1 above.

[0144] Positive drug 1: Protoporphyrin Positive drug 2: 5-aminolevulinic acid Solvent: DMSO.

[0145] 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 1010 CFU / mL (1 OD≈9.52*10 9 CFU / mL) for later use.

[0146] 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.

[0147] 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.

[0148] 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.

[0149] 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.

[0150] The antibacterial rate calculation formula is as follows:

[0151] 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.

[0152] 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.

[0153] 2. Results and Discussion 2.1 Results of the first biological replication The results are as follows Figure 6 As shown in (a), compared with the blank illumination group, the optical density of compound N-02 and the optical density of positive drug 1 were significantly reduced (p<0.0001), and their inhibition rates were 44.53% and 32.65%, respectively.

[0154] 2.2 Results of the second biological replication The results are as follows Figure 6 As shown in (b), compared with the blank illumination group, the optical density of compound N-02 and positive drug 1 was significantly reduced (p<0.0001). The inhibition rates of N-02 and positive drug 1 were 47.4% and 28.2%, respectively.

[0155] 2.3 Results of the third biological replication The results are as follows Figure 6 As shown in (c), compared with the blank illumination group, the optical density of compound N-02 and positive drug 1 was significantly reduced (p<0.0001). The inhibition rates of N-02 and positive drug 1 were 46.8% and 32.4%, respectively.

[0156] 2.4 Inhibition rate of three biological replicates The results are as follows Figure 6 As shown in (d) and Table 5, the compound N-02 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).

[0157] Table 5: Inhibitory effect of compounds N-02, PPIX and 5-ALA on Porphyromonas gingivalis .

Claims

1. A compound of formula (I) or a salt thereof: (I) in: R stands for -NR 1 R 2 , R 1 and R 2 Each independently represents hydrogen, 3-10 membered heterocyclyl (C1-C8)-alkyl or 5-12 membered heteroaryl (C1-C8)-alkyl, wherein R 1 and R 2 Different from hydrogen, wherein the heterocyclic group or heteroaryl group contains one or two of the heteroatoms of O, S and N; The condition is to exclude compounds and .

2. The compound or salt thereof according to claim 1, wherein R 1 and R 2 Each independently represents hydrogen, 3-8 membered heterocyclyl(C1-C6)-alkyl or 5-10 membered heteroaryl(C1-C6)-alkyl.

3. The compound or salt thereof according to claim 2, wherein R 1 and R 2 Each independently represents hydrogen, morpholino(C1-C6)-alkyl or thiophene(C1-C6)-alkyl.

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 in the preparation of a medicament for the treatment of a hyperproliferative disease.

8. The use according to claim 7, wherein the hyperproliferative disease is cancer or a precancerous lesion.

9. The method of claim 8, wherein the cancer is selected from bladder cancer, esophageal cancer, bronchial cancer, oral cancer, nasopharyngeal cancer, liver cancer, pancreatic cancer, skin cancer, penile cancer, cervical cancer, vaginal cancer, endometrial cancer, ovarian cancer, colorectal cancer, kidney cancer, urothelial cancer, thyroid cancer, breast cancer, anal cancer, Kaposi's sarcoma, lung cancer, gastric cancer, bile duct cancer, prostate cancer, melanoma or brain cancer; The precancerous lesions are selected from cervical precancerous lesions, oral leukoplakia, myelodysplastic diseases, familial intestinal polyps, skin nevus, psoriasis or solar keratosis.

10. 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.

11. The method according to claim 10, 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.

12. The use according to claim 10, 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.

13. 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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