Use of mitochondrial division inhibitors and combinations comprising the same

Abstract

The present application relates to the field of biomedicine. Specifically, the present application provides the use of a mitochondrial fission inhibitor in the preparation of a medicament for preventing or treating opioid analgesic dependence or enhancing the analgesic effect of an opioid analgesic. The present application also provides a combination comprising a mitochondrial fission inhibitor and the use thereof.

Description

[0001] This application claims priority to Chinese Patent Application No. 202011410107.5, filed on December 3, 2020, entitled “Use of a mitochondrial division inhibitor and a combination thereof”, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0002] This invention relates to the field of biomedical technology. Specifically, this invention provides the use of mitochondrial fission inhibitors in the preparation of medicaments for the prevention or treatment of opioid analgesia dependence or for enhancing the analgesic effect of opioid analgesics. This invention also provides combinations comprising mitochondrial fission inhibitors and their uses. Background Technology

[0003] Opioid analgesics are commonly used analgesics in clinical practice, with morphine being one of the top five most abused / used medical drugs. Long-term use of opioid analgesics can easily lead to drug dependence, causing patients to develop physiological tolerance to the drugs, thus reducing their analgesic effect, and inducing physical and psychological withdrawal symptoms upon discontinuation.

[0004] Furthermore, long-term use of opioid analgesics can cause adaptive changes in brain function. Disruptions in neuronal energy homeostasis leading to neuronal dysfunction are a significant cause of the development and progression of neurodegenerative diseases (such as Parkinson's and Alzheimer's) and mental illnesses (such as schizophrenia, bipolar disorder, and depression).

[0005] Mitochondria, as energy-producing organelles in eukaryotic cells, efficiently synthesize ATP to meet cellular energy metabolism needs, while also regulating cellular oxidative stress, signal transduction, apoptosis, calcium homeostasis, and neural plasticity. Mitochondria are highly dynamic organelles, and their fusion and division are crucial for the regulation of mitochondrial function. Current research suggests that mitochondrial fragmentation (i.e., loss of mitochondrial fusion function) can impair cellular function, such as causing damage to mitochondrial respiration and loss of membrane potential.

[0006] Dynamin-related protein 1 (Drp1) is a key protein regulating mitochondrial division. Mdivi-1 (see, for example, WO 2005 / 051974 and WO 2019 / 126179) is a selective inhibitor of Drp1 that inhibits mitochondrial division. Mdivi-1 has been reported to have cytoprotective effects in neuronal degenerative diseases such as Parkinson's disease and myocardial damage (see, for example, Bido, S et al., Sci Rep., 2017, 7(1):7495; Ong, SB et al., Int JMol Sci., 2019, 20(16):3972). The short peptides DLLPRGT or DLLPRGS in human Drp1 (see, for example, WO2012 / 158624) can inhibit mitochondrial division by specifically inhibiting the GTPase activity of Drp1. Summary of the Invention

[0007] In a general aspect, the present invention provides the use of a mitochondrial splitting inhibitor in the preparation of a medicament for the prevention or treatment of opioid analgesia dependence or for enhancing the analgesic effect of opioid analgesics. In some embodiments, the mitochondrial splitting inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof. In other embodiments, the mitochondrial splitting inhibitor is a compound of formula (III) or a pharmaceutically acceptable salt thereof.

[0008] In some embodiments, the mitochondrial division inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound of formula (I) has the following structure:

[0009]

[0010] Where R 1 Selected from: hydrogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 cycloalkyl, C 1-6 Heteroalkyl, C 2-6 Heterene, C 2-6 heteroyne group and C 3-8 Heterocyclic alkyl groups;

[0011] R 2 R 3 R 4 and R 5 Each is independently selected from: hydrogen, C 1-6 Alkyl, C 1-6 Heteroalkyl groups and halogens;

[0012] The alkyl, alkenyl, ynyl, cycloalkyl, heteroalkyl, heteroalkenyl, heteroynyl, heterocyclic, and alkoxy groups thereon are optionally substituted with one or more of the following groups: halogen, C 1-6 Alkyl, cyano, C 1-6 Alkoxy, C 1-6 Alkylthio, hydroxyl, SH, SO4, C 3-8 cycloalkyl, C 3-8 Heterocyclic alkyl, amino, amide, oxo (O=) and nitro groups.

[0013] In one specific embodiment, the compound of formula (I) has the structure of formula (II):

[0014]

[0015] In other embodiments, the mitochondrial division inhibitor is a compound of formula (III) or a pharmaceutically acceptable salt thereof, wherein the compound of formula (III) has the following structure:

[0016] (CP1-Linker1) p -Z-(Linker2-CP2) q (III)

[0017] in

[0018] CP1 and CP2 are each independently transmembrane peptides;

[0019] Linker1 and Linker2 either do not exist independently, or they are connectors;

[0020] Z is a mitochondrial division inhibitory peptide;

[0021] p and q are each independently 0 or 1;

[0022] Preferably, p is 0 and q is 0 or 1; or p is 1 and q is 0.

[0023] In some implementations, Z is [D / E]-[L / I / M / V / F]-[L / I / M / V / F]-P-[R / K]-[G / A / T / S]-[T / S / A / G]. In some implementations, Z is DLLPRG-[T / S / A / G]. In some implementations, Z is DLLPRG-[S / T].

[0024] In some embodiments, Z is X1-[D / E]-[L / I / M / V / F]-[L / I / M / V / F]-P-[R / K]-[G / A / T / S]-[T / S / A / G]-X2, wherein X1 and X2 may exist simultaneously or not simultaneously, X1 is selected from [G / A / S]-[R / K], R, and K, and X2 is selected from [G / A / S]-[I / L / V], G, A, and S. In some embodiments, Z is X1-DLLPRG-[T / S / A / G]-X2. In some embodiments, Z is X1-DLLPRG-[S / T]-X2.

[0025] In some embodiments, the membrane-penetrating peptide comprises an amino acid sequence selected from SEQ ID NO:3-SEQ ID NO:14 or other amino acid sequences, said other amino acid sequences comprising 3-50 consecutive or discontinuous arginine residues. In one specific embodiment, the membrane-penetrating peptide comprises an amino acid sequence selected from SEQ ID NO:3-SEQ ID NO:6. In a preferred embodiment, the membrane-penetrating peptide comprises an amino acid sequence of SEQ ID NO:3 or SEQ ID NO:5.

[0026] In some embodiments, the linker comprises 1-20 amino acids. In some embodiments, the linker is selected from (G). n G n A, (GS) n G n S m 、(G n S) m G n S, where n and m are each independently selected from integers from 1 to 20. In some preferred embodiments, the connector is selected from G, GG, GGG, GGGG, GGSG, GGGGS, GGGSG, and GSSSG. In one specific embodiment, the connector is GG.

[0027] In one specific embodiment, the compound of formula (III) is [D / E]-[L / I / M / V / F]-[L / I / M / V / F]-P-[R / K]-[G / A / T / S]-[T / S / A / G]. In one specific embodiment, the compound of formula (III) is DLLPRG-[T / S / A / G]. In one specific embodiment, the compound of formula (III) has the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2.

[0028] In one embodiment, the compound of formula (III) has an amino acid sequence selected from SEQ ID NO:15-SEQ ID NO:18. In another embodiment, the compound of formula (III) has an amino acid sequence of SEQ ID NO:15 or SEQ ID NO:17.

[0029] In another aspect, the present invention provides a combination comprising: (1) a mitochondrial division inhibitor as described above; and (2) an opioid analgesic; optionally, the combination further comprises a non-opioid analgesic and / or an adjunct analgesic. In some embodiments, the opioid analgesic is selected from morphine, dihydromorphine, diacetylmorphine, hydromorphone, hydroxymorphone, oxycodone, hydrocodone, codeine, dihydrocodeine, buprenorphine, methadone, fentanyl, remifentanil, sufentanil, buprenorphine, meperidine, dezocine, butorphanol, bremazocine, carfentanyl, cyclazocine, enadoline, eseroline, ethylmorphine, etonitazine, etorphanine, levofenphene, levofenol, nalbuphine, nicomorphine, pentazocine, phenazocine, picenadol, tramadol, tapentadol, pharmaceutically acceptable salts thereof, and combinations thereof. In some embodiments, the mitochondrial division inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, the mitochondrial division inhibitor is Mdivi-1 or a pharmaceutically acceptable salt thereof. In other embodiments, the mitochondrial division inhibitor is a compound of formula (III) or a pharmaceutically acceptable salt thereof. In some embodiments, the mitochondrial division inhibitor has the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2. In some embodiments, the mitochondrial division inhibitor has an amino acid sequence selected from SEQ ID NO:15-SEQ ID NO:18.

[0030] In another aspect, the present invention also provides the use of the said combination in the preparation of a medicament for providing analgesia to a subject. In some embodiments, the medicament is used to treat chronic pain, acute pain, postoperative pain, intermittent pain, penetrating pain, tearing pain, mild pain, moderate pain, severe pain, excruciating pain, bone pain, joint pain, soft tissue pain, neuralgia, cancer pain, inflammatory pain, and traumatic pain; preferably, the medicament is used to treat cancer pain, chronic pain, and inflammatory pain; more preferably, the medicament is used to treat moderate or severe pain. Attached Figure Description

[0031] Figure 1ALong-term morphine use leads to increased mitochondrial fragmentation in dopaminergic neurons in the ventral tegmental area of ​​the midbrain. The mitochondrial area in the dendrites of VTA dopaminergic neurons in mice in the saline-treated (Sal) and morphine-treated (Mor) groups is shown; each point represents the area (μm) of a single mitochondria in the dendrites of a dopaminergic neuron. 2 Left: Bar chart; Right: Cumulative probability distribution curve; *P<0.05, ****P<0.0001.

[0032] Figure 1B Effects of Mdivi-1 on mitochondrial fragmentation of dopaminergic neurons in the ventral tegmental area of ​​the midbrain induced by long-term morphine use. Mitochondrial area in dendrites of VTA dopaminergic neurons in mice subjected to escalating doses of morphine (Mdivi-1 group) and the control group (Vehicle); each point represents the area (μm) of a single mitochondria in the dendrites of a dopaminergic neuron. 2 Left: Bar chart; Right: Cumulative probability distribution curve; *P<0.05, ****P<0.0001.

[0033] Figure 1C Effects of P110 on mitochondrial fragmentation of dopaminergic neurons in the ventral tegmental area of ​​the midbrain induced by long-term morphine use. The P110 group (P110) and the control protein group (TAT) underwent escalating morphine administration. (47-57) ) Mitochondrial area in dendrites of VTA dopaminergic neurons in mice; each point represents the mitochondrial area (μm) in the dendrites of a single dopaminergic neuron. 2 Left: Bar chart; Right: Cumulative probability distribution curve; ****P<0.0001.

[0034] Figure 2A Effect of Mdivi-1 on analgesic tolerance induced by long-term morphine use. The percentage of maximum analgesic effect (MPE%) over time was plotted in control mice (Mor+vehicle) and Mdivi-1 groups (Mor+Mdivi-1 (12.5 mg / kg), Mor+Mdivi-1 (25 mg / kg), Mor+Mdivi-1 (50 mg / kg), Mor+Mdivi-1 (100 mg / kg)) after subcutaneous morphine administration (10 mg / kg once daily for 6 consecutive days). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0035] Figure 2B Effect of P110 on analgesic tolerance induced by long-term morphine use. The control group (Mor+TAT) experienced subcutaneous morphine administration (10 mg / kg once daily for 6 consecutive days). (47-57)Maximum analgesic percentage (MPE%) plotted over time (days) in mice in groups P110 (Mor+P110 (0.2 mg / kg), Mor+P110 (0.5 mg / kg), and Mor+P110 (1 mg / kg); **P<0.01, ***P<0.001, ****P<0.0001.

[0036] Figure 3 Effects of Mdivi-1 on somatic symptoms induced by naloxone-induced acute morphine withdrawal. The somatic symptoms of naloxone-induced acute morphine withdrawal in control (Vehicle) and Mdivi-1 group (Mdivi-1) mice were analyzed. Each point represents the number of occurrences of each symptom (from left to right: wet dog shakes, body tremor, backward locomotion, and piloerection) in a single mouse within 20 minutes; **P<0.01, ***P<0.001, ****P<0.0001.

[0037] Figure 4 Effects of Mdivi-1 on morphine withdrawal-induced discomfort symptoms. Six days after spontaneous morphine withdrawal, the CPA scores of mice in the control group (Vehicle) and the Mdivi-1 group (Mdivi-1) underwent conditioned place aversion (CPA) training were analyzed; CPA score (seconds) = time on the restricted side (seconds) - time on the untrained side (seconds); Pre-test, before CPA training; Test, after CPA training; **P<0.01.

[0038] Figure 5 Effect of Mdivi-1 on anxiety levels induced by spontaneous morphine withdrawal. Seven days after spontaneous morphine withdrawal, anxiety levels in the control group (Vehicle) and the Mdivi-1 group (Mdivi-1) were analyzed using the high cross maze test. Each point represents the time (in seconds) a single mouse spent in the open arm of the high cross maze. ***P<0.001.

[0039] Figure 6 Effect of Mdivi-1 on depression levels induced by spontaneous morphine withdrawal. Eight days after spontaneous morphine withdrawal, depression levels in the control group (Vehicle) and the Mdivi-1 group (Mdivi-1) mice were analyzed using the suspended tail test (TST). Each point represents the time (in seconds) that a single mouse remained still within 6 minutes; **P<0.01.

[0040] Figure 7The effect of Mdivi-1 on social avoidance behavior induced by spontaneous morphine withdrawal. Nine days after spontaneous morphine withdrawal, the social avoidance levels of mice in the control group (Vehicle) and the Mdivi-1 group (Mdivi-1) were analyzed using a three-box social test. Each point represents the social preference index of a single test mouse, calculated as: social preference index = time spent interacting with a stranger mouse / time spent exploring an empty cage; **P<0.01. Detailed Implementation

[0041] definition

[0042] In this invention, unless otherwise stated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. All referenced literature, patents, and patent applications are incorporated herein by reference in their entirety. Furthermore, the terms and laboratory procedures related to protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, and immunology used herein are widely used terms and routine procedures in their respective fields. To better understand this invention, definitions and explanations of relevant terms are provided below.

[0043] Unless otherwise stated, the term "combination thereof" refers to a multi-component mixture of the elements, such as two, three, four, and up to the maximum possible multi-component mixture.

[0044] In this document, if the number of elements or components is not specified beforehand, it means that there is no limit to the number of occurrences (or presence) of the elements or components. Therefore, it should be interpreted as including one or at least one, and the singular form of a component or component also includes the plural, unless the value clearly indicates a singular.

[0045] As used herein, “at least one species” or “one or more species” can mean 1, 2, 3, 4, 5, 6, 7, 8 or more species.

[0046] In this document, when referring to a range, preferred range, or preferred upper and lower limits of a parameter (e.g., a therapeutically effective dose), it should be understood that this is equivalent to specifically disclosing any range by combining any pair of upper or preferred values ​​with any lower or preferred value, regardless of whether the range or the value is specifically disclosed. Unless otherwise stated, the numerical ranges listed herein are intended to include the endpoints of the range and all integers and fractions (decimals) within that range.

[0047] As used herein, the terms “about” or “approximately” when used with a numerical variable generally mean that the value of the variable and all values ​​of the variable are within the experimental error (e.g., within a 95% confidence interval for the mean) or within ±10% of the specified value, or a wider range.

[0048] In this document, the expressions "comprising," or similar synonyms such as "including," "containing," and "having," are open-ended and do not exclude additional unlisted elements, steps, or components. The expression "consisting of..." excludes any unspecified elements, steps, or components. The expression "substantially consisting of..." limits the scope to the specified elements, steps, or components, plus optional elements, steps, or components that do not materially affect the essential and novel features of the claimed subject matter. It should be understood that the expression "comprising" encompasses both the expressions "substantially consisting of..." and "consisting of...".

[0049] As used herein, the terms “optional” or “optionally present” mean that an event or condition subsequently described may or may not occur, including both the occurrence and non-occurrence of said event or condition. For example, when a group is described as optionally substituted, it may be unsubstituted or substituted, for example, substituted by one or more substituents independently selected from those described herein. Those skilled in the art will understand that “optional” also encompasses the meaning that the type and number of substituents can be arbitrarily chosen and combined, provided that the resulting compound is stable.

[0050] As used herein, the term "substituted" means that one or more hydrogen atoms of a specified atom are selectively replaced by a given group, provided that the substitution does not exceed the normal valence of the specified atom in the present case and that the substitution forms a stable compound. Combinations of substituents and / or variables are permitted only if such combinations form a stable compound. Examples of substituents for the optionally substituted group referred to herein may be halogens, C... 1-6 Alkyl, cyano, C 1-6 Alkoxy, C 1-6 Alkylthio, hydroxyl, SH, C 3-8 cycloalkyl, C 3-8 One or more of heterocyclic alkyl, SO4, amino, amide, oxo (O=) and nitro groups.

[0051] As used in this article, the term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0052] In this paper, "ij" or similar expressions refer to a range that encompasses the endpoints, each integer within it, and the subrange consisting of those integers. For example, C 1-6 Covering C1, C2, C3, C4, C5, and C6, as well as C 2- C5, C 3- C4, etc.

[0053] As used herein, the term "alkyl," whether used alone or in combination with other terms, refers to a branched or straight-chain saturated aliphatic hydrocarbon group having a specific number of carbon atoms. Unless otherwise specified, "alkyl" refers to a C14-C14 group. 1-6 Alkyl group. For example, "C 1-6 "Alkyl" includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, hexyl, etc. The alkyl groups described herein may optionally be substituted.

[0054] As used herein, the term "alkenyl," whether used alone or in combination with other terms, refers to a branched or straight-chain unsaturated aliphatic hydrocarbon group containing a specific number of carbon atoms and a carbon-carbon double bond. Unless otherwise noted, "alkenyl" refers to a C14-carbon group. 2-6 Alkenyl group. The alkenyl group described herein may optionally be substituted.

[0055] As used herein, the term "alkynyl" (whether used alone or in combination with other terms) refers to a branched or straight-chain unsaturated aliphatic hydrocarbon group containing a specific number of carbon atoms and a carbon-carbon triple bond. Unless otherwise noted, "alkynyl" refers to a C-carbon group. 2-6 Alkyne group. The alkynyl group described herein may optionally be substituted.

[0056] As used in this article, "heteroatoms" include nitrogen, oxygen, and sulfur.

[0057] As used herein, the term "heteroalkyl" (whether used alone or in combination with other terms) refers to the alkyl group described above, which is attached to other parts of the molecule by a heteroatom. Examples include, but are not limited to, alkoxy and alkathiol groups. In a preferred embodiment, the heteroalkyl group is C10. 1-6 Alkoxy or C 1-6 Alkylthio group. The heteroalkyl groups described herein may optionally be substituted.

[0058] As used herein, the term "alkoxy," whether used alone or in combination with other terms, refers to the alkyl group described above, which is attached to other parts of the molecule by an oxygen atom (alkyl-O-). Unless otherwise noted, "alkoxy" refers to C 1-6 Alkyl groups. For example, "C 1-6 "Alkoxy" includes, but is not limited to, methoxy (C1 alkoxy), ethoxy (C2 alkoxy), propoxy (C3 alkoxy), butoxy (C4 alkoxy), etc. The alkoxy groups described herein may optionally be substituted.

[0059] As used herein, the term "alkylthio," whether used alone or in combination with other terms, refers to the alkyl group described above, which is attached to other parts of the molecule by a sulfur atom (alkyl-O-). Unless otherwise noted, "alkylthio" refers to C 1-6 Alkylthio group. For example, C 1-6Alkylthio groups include, but are not limited to, methylthio (C1 alkylthio), ethylthio (C2 alkylthio), propylthio (C3 alkylthio), and butylthio (C4 alkylthio).

[0060] As used herein, the term "heteroalkenyl," whether used alone or in combination with other terms, refers to the alkenyl group described above, which is attached to other parts of the molecule by a heteroatom. The heteroalkenyl group described herein may optionally be substituted.

[0061] As used herein, the term "heterynyl" (whether used alone or in combination with other terms) refers to the ynyl group described above, which is attached to other parts of the molecule via a heteroatom. The heterynyl group described herein may optionally be substituted.

[0062] As used herein, the term "cycloalkyl," whether used alone or in combination with other terms, refers to a saturated aliphatic cyclic hydrocarbon group having a specific number of carbon atoms. Unless otherwise noted, "cycloalkyl" refers to a C14-C14 cyclic hydrocarbon group. 3-8 Cycloalkyl groups. For example, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. The cycloalkyl groups described herein may optionally be substituted.

[0063] As used herein, the term "heterocyclic alkyl," whether used alone or in combination with other terms, refers to a monocyclic or polycyclic alkyl ring in which one, two, or three of the carbide ring atoms are replaced by heteroatoms selected from N, O, and S. Unless otherwise specified, "heterocyclic alkyl" refers to a C14-C14 ring. 3-8 Heterocyclic alkyl groups. Examples include, but are not limited to, piperazinyl, piperidinyl, tetrahydropyranyl, tetrahydrofuranyl, 1,3-dioxanehexyl, etc. The heterocyclic alkyl groups described herein may optionally be substituted.

[0064] As used herein, the term "peptide" or "polypeptide" refers to a compound containing two or more covalently linked amino acids. Unless otherwise specified, "peptide" or "polypeptide" may be used interchangeably. In this document, the sequence of a polypeptide is represented by a single-letter amino acid from the N-terminus to the C-terminus. For example, GR indicates that the carboxyl group of glycine (G) and the amino group of arginine (R) form a peptide bond.

[0065] Typically, one or more (e.g., 1, 2, 3, 4, 5, 6, 7) amino acids in a polypeptide (e.g., mitochondrial fission inhibitory peptides or membrane-penetrating peptides) can be substituted with suitable conserved amino acids without generally altering its biological activity (see, for example, Watson et al., Molecular Biology of the Gene, 4th Edition, 1987, The Benjamin / Cummings Pub.co., p. 224). Suitable conserved substitutions are well known to those skilled in the art. Some common non-limiting examples of conserved amino acid residue substitutions are listed in the table below.

[0066] Ala(A) Val, Leu, Ile Val Arg(R) Lys, Gln, Asn Lys Asn(N) Gln, His, Asp, Lys, Arg Gln Asp(D) Glu, Asn Glu Cys(C) Ser, Ala Ser Gln(Q) Asn, Glu Asn Glu(E) Asp, Gln Asp Gly(G) Ala,Thr,Ser Ala His(H) Asn, Gln, Lys, Arg Arg Ile(I) Leu, Val, Met, Ala, Phe Leu Leu(L) Ile, Val, Met, Ala, Phe Ile Lys(K) Arg, Gln, Asn Arg Met(M) Leu, Phe, Ile Leu Phe(F) Trp, Leu, Val, Ile, Ala, Tyr Tyr Pro(P) Ala Ala Ser(S) Thr Thr Thr(T) Ser, Gly, Ala, Val Ser Trp(W) Tyr, Phe Tyr Tyr(Y) Trp, Phe, Thr, Ser Phe Val(V) Ile, Leu, Met, Phe, Ala Leu

[0067] As used in this article, the expression [A / B / C] indicates that the variable at that position can be selected from A, B, and C. For example, [L / I / M / V / F] indicates that the amino acid at that position can be selected from L, I, M, V, and F.

[0068] As used herein, the term "pharmaceutically acceptable" means a substance that, within the bounds of normal medical judgment, is suitable for contact with the tissues of a subject without undue toxicity, irritation, allergic reactions, etc., has a reasonable risk-benefit ratio, and is effective for its intended use.

[0069] The term "pharmaceutically acceptable salt" refers to salts made from pharmaceutically acceptable, non-toxic bases or acids, including inorganic or organic bases and inorganic or organic acids. Salts of inorganic bases can be selected from, for example, salts of aluminum, ammonium, calcium, copper, iron, ferrous, lithium, magnesium, manganese, divalent manganese, potassium, sodium, and zinc. Salts of organic bases can be, for example, salts formed with substances such as N-methylglucosamine, dimethylglucosamine, ethylglucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, sarcosine, serine, trihydroxymethylaminomethane, and aminopropylene glycol, etc. Salts of inorganic or organic acids can be, for example, acid addition salts formed with acids selected from: acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, viscous acid, nitric acid, pyric acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, and p-toluenesulfonic acid. Those skilled in the art will further recognize that the acid addition salts of the compounds of the present invention can be prepared by reacting the compounds with a suitable inorganic or organic acid using any of a variety of known methods. Alternatively, alkali metal salts and alkaline earth metal salts of the acidic compounds of the present invention can be prepared by reacting the compounds of the present invention with a suitable base using various known methods.

[0070] As used herein, a "pharmaceutically acceptable carrier" may include, but is not limited to, diluents, buffers, dispersants, glidants, sweeteners, solubilizers, antioxidants, absorption delayers, disintegrants, binders, lubricants, preservatives, emulsifiers, surfactants, solvents, softeners, chelating agents, gelling agents, adjuvants, colorants, flavorings, humectants, osmotic pressure regulators, coatings, and structure-forming excipients. Exemplary pharmaceutically acceptable carriers may include, for example, trehalose, sorbitol, lactose, glucose, dextrin, starch, inorganic salts, mannitol, cellulose, gelatin, syrups, polysorbate 80, polysorbate 20, poloxamer, ascorbic acid, citric acid, cysteine ​​hydrochloride, talc, distilled water, ethanol, and vegetable oils.

[0071] As used herein, "mitochondrial fission inhibitor" refers to a substance capable of inhibiting mitochondrial fission, thereby inhibiting mitochondrial fragmentation and maintaining the elongated network structure of mitochondria. Generally, mitochondrial fission inhibitors target mitochondrial fission-related proteins (e.g., Drp1, Fis1 / Mdv2, and Fis2 / Mdv1) and inhibit mitochondrial fission by inhibiting these proteins (e.g., inhibiting the activity, function, expression, or protein-protein interactions of Drp1, Fis1 / Mdv2, and Fis2 / Mdv1). For example, Mdivi-1 has been reported to inhibit mitochondrial fission by inhibiting Drp1 self-assembly and GTPase activity. Mitochondrial fission inhibitors that can be used in this invention include, but are not limited to, small molecule compounds, peptides, or salts thereof.

[0072] As used herein, “opioid analgesics” refers to substances that act on opioid receptors and produce an analgesic effect. Opioid receptors may include, for example, μ, δ, κ, and σ opioid receptors. Opioid analgesics may be natural or synthetic. Opioid analgesics may include, but are not limited to, opioid receptor agonists (e.g., morphine, oxycodone, methadone, and fentanyl), partial agonists (e.g., buprenorphine), and agonist-antagonists (e.g., dezocine and pentazocine). Opioid analgesics may be compounds having a single active ingredient or mixtures containing multiple active ingredients (analgesic activities). Exemplary opioid analgesics may include, but are not limited to: morphine, dihydromorphine, diacetylmorphine, hydromorphone, oxymorphone, oxycodone, hydrocodone, codeine, dihydrocodeine, bucinnazine, methadone, fentanyl, remifentanil, sufentanil, buprenorphine, pethidine, and dezocine. ocine), butorphanol, bremazocine, carfentanyl, cyclazocine, enadoline, eseroline, ethylmorphine, etonitazine, etorphine, levophenacylmorphan, levorphanol, nalbuphine, nicomorphine, pentazocine, phenazocine, picenadol, tramadol, tapentadol, and their pharmaceutically acceptable salts.

[0073] In this article, "pain" may include, but is not limited to, chronic pain, acute pain, postoperative pain, intermittent pain, penetrating pain, tearing pain, mild pain, moderate pain, severe pain, extreme pain, bone pain, joint pain, soft tissue pain, neuralgia, disease pain (such as cancer pain, inflammatory pain), and traumatic pain.

[0074] In this document, the terms "treatment of pain" or "providation of analgesia" can include the prevention, reduction, or elimination of pain. In some embodiments, analgesia is provided to a subject by administering a combination of Mdivi-1 and morphine. In some embodiments, analgesia is provided to a subject by administering a combination of P110 and morphine.

[0075] The compounds referred to herein, such as mitochondrial fission inhibitors and opioid analgesics, also encompass their salts, esters, solvates (such as hydrates), prodrugs, formulations, crystal forms, cocrystals, stereoisomers, tautomers, and isotopic compounds. Preferably, these forms are pharmaceutically acceptable.

[0076] As used herein, "therapeutic effective amount" means an amount that, when administered to a subject, is at least sufficient to produce a therapeutic effect, such as an amount sufficient to prevent and treat opioid dependence, or an amount sufficient to produce an analgesic effect. Therapeutic effective amounts may be administered in one or more doses. Those skilled in the art can determine the therapeutic effective amount based on the size of the subject, the severity of symptoms, and the specific combination of drugs chosen or the route of administration.

[0077] As used herein, the term "subject" refers to mammals, including but not limited to humans and non-human primates, cattle, pigs, horses, sheep, cats, dogs, rats, and mice, particularly humans. In some embodiments, the subject is a pain patient. In some embodiments, the subject is a person who requires long-term use of opioid analgesics to treat pain, such as a patient with cancer, inflammation, or other serious illness.

[0078] I. Uses of mitochondrial division inhibitors

[0079] In one general aspect, the present invention provides the use of mitochondrial fission inhibitors in the preparation of medicaments for the prevention or treatment of opioid analgesia dependence or for enhancing the analgesic effect of opioid analgesics.

[0080] In another aspect, the present invention relates to mitochondrial fission inhibitors for the prevention or treatment of opioid dependence or to enhance the analgesic effect of opioid analgesics.

[0081] 1. Inhibitors of mitochondrial division

[0082] Mitochondrial division inhibitors may include, but are not limited to, compounds of formula (I) and their pharmaceutically acceptable salts, compounds of formula (III) and their pharmaceutically acceptable salts, and combinations thereof.

[0083] In some embodiments, the mitochondrial division inhibitor is selected from compounds of formula (I) and their pharmaceutically acceptable salts, compounds of formula (III) and their pharmaceutically acceptable salts, and combinations thereof.

[0084] 1.1 Compounds of formula (Ⅰ) and their pharmaceutically acceptable salts

[0085] In some embodiments, the mitochondrial division inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound of formula (I) has the following structure:

[0086]

[0087] Where R 1 Selected from: hydrogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 cycloalkyl, C 1-6 Heteroalkyl, C 2-6 Heterene, C 2-6 heteroyne group and C 3-8 Heterocyclic alkyl groups;

[0088] R 2 R 3 R 4 and R 5 Each is independently selected from: hydrogen, C 1-6 Alkyl, C 1-6 Heteroalkyl groups and halogens;

[0089] The alkyl, alkenyl, ynyl, cycloalkyl, heteroalkyl, heteroalkenyl, heteroynyl, and heterocycloalkyl groups are optionally substituted with one or more of the following groups: halogen, C 1-6 Alkyl, cyano, C 1-6 Alkoxy, C 1-6 Alkylthio, hydroxyl, SH, SO4, C 3-8 cycloalkyl, C 3-8 Heterocyclic alkyl, amino, amide, oxo (O=) and nitro.

[0090] In one implementation scheme, R 4 and R 5 Each is an independent halogen.

[0091] In one implementation scheme, R 3 Selected from C 1-6 Heteroalkyl groups.

[0092] In one implementation scheme, R 3 It is a C1 alkoxy group.

[0093] In one implementation scheme, R 3 It is a C3 alkoxy group.

[0094] In one implementation scheme, R 3 It is a C1 alkoxy group, R 2 It is hydrogen.

[0095] In one implementation scheme, R 3 It is a C3 alkoxy group, R 2 It is hydrogen.

[0096] In one implementation scheme, R 3 It is a C3 alkoxy group, R 1 Independently selected from hydrogen and C 1-6 alkyl.

[0097] In one implementation scheme, R 3 It is a C1 alkoxy group, R 2 For hydrogen, R 1 It is hydrogen.

[0098] In one implementation scheme, R 3 It is a C1 alkoxy group, R 2 For hydrogen, R 1 It is a C1 alkyl group.

[0099] In one implementation scheme, R 3 It is a C3 alkoxy group, R 2 For hydrogen, R 1 It is hydrogen.

[0100] In one implementation scheme, R 3 It is a C1 alkoxy group, R 2 The compound of formula (I) is hydrogen and has the structure of formula (II) (also known as Mdivi-1):

[0101]

[0102] Descriptions and preparation methods of compounds of formula (I) and their pharmaceutically acceptable salts can be found, for example, in WO2005 / 051974 and WO 2019 / 126179 and CN 111329864A, the contents of which are incorporated herein by reference in their entirety. In some embodiments, Mdivi-1 is commercially available.

[0103] 1.2 Compounds of formula (III) and their pharmaceutically acceptable salts

[0104] In some embodiments, the mitochondrial division inhibitor is a compound of formula (III) or a pharmaceutically acceptable salt thereof, wherein the compound of formula (III) has the following structure:

[0105] (CP1-Linker1) p -Z-(Linker2-CP2) q (III)

[0106] in

[0107] CP1 and CP2 are each independently transmembrane peptides;

[0108] Linker1 and Linker2 either do not exist independently, or they are connectors;

[0109] Z is a mitochondrial division inhibitory peptide;

[0110] p and q are each independently 0 or 1, that is, CP1-Linker1 and Linker2-CP2 exist or do not exist independently; preferably, p is 0 and q is 0 or 1; or p is 1 and q is 0.

[0111] Mitochondrial division inhibitory peptide

[0112] In this document, "mitochondrial division inhibitory peptide" refers to a peptide capable of inhibiting mitochondrial division. There are no limitations on the mitochondrial division inhibitory peptides that can be used in this invention, as long as they possess the activity of inhibiting mitochondrial division.

[0113] The preferred mitochondrial division inhibitory peptide comprises the amino acid sequence of SEQ ID NO:1 (DLLPRGT) or a variant thereof. For a description of SEQ ID NO:1 or a variant thereof, see, for example, WO 2012 / 158624, the entire contents of which are incorporated herein by reference. SEQ ID NO:1 is amino acids 49-55 of the human Drp1 protein (Uniprot identification number: O00429-1).

[0114] Variants of SEQ ID NO:1 can be obtained by using conservative amino acid substitutions for amino acids 1, 2, 3, 4, 5, 6, or 7 in SEQ ID NO:1. Conservative amino acid substitutions include, for example, replacing aspartic acid (D) at position 1 with glutamic acid (E), leucine (L) at position 2 or 3 with isoleucine (I), methionine (M), valine (V), or phenylalanine (F), arginine (R) at position 5 with lysine (L), glycine (G) at position 6 with alanine (A), threonine (T), or serine (S), and threonine (T) at position 7 with glycine (G), alanine (A), or serine (S). An example of a variant of SEQ ID NO:1 obtained by conservative amino acid substitutions is, for example, SEQ ID NO:2.

[0115] In some implementations, the mitochondrial division inhibitory peptide has the structure of formula (Ⅳ):

[0116] aa1-aa2-aa3-aa4-aa5-aa6-aa7(IV)

[0117] Where aa1 is D or E; aa2 and aa3 are each independently selected from L, I, M, V and F; aa4 is P; aa5 is R or K; aa6 is selected from G, A, T and S; and aa7 is selected from S, T, G and A.

[0118] In some embodiments of formula (Ⅳ), the mitochondrial mitochondrial mitochondrial inhibitory peptide is [D / E]-[L / I / M / V / F]-[L / I / M / V / F]-P-[R / K]-[G / A / T / S]-[T / S / A / G]. In other embodiments, aa1 is D, aa2 and aa3 are L, aa4 is P, aa5 is R, aa6 is G, and aa7 is selected from T, S, A, and G, and the mitochondrial mitochondrial inhibitory peptide is DLLPRG-[T / S / A / G]. In one specific embodiment, the mitochondrial mitochondrial inhibitory peptide has the amino acid sequence of SEQ ID NO:1 (DLLPRGT) or SEQ ID NO:2 (DLLPRGS).

[0119] Variants of SEQ ID NO:1 can also be obtained by adding 1-2 amino acids to the N-terminus and / or C-terminus of SEQ ID NO:1. For example, amino acids that can be added to the N-terminus include GR and R. The G can be replaced with A or S. The R can be replaced with K. For example, amino acids that can be added to the C-terminus include GI and G. The G can be replaced with A or S. The I can be replaced with L or V. Such variants of SEQ ID NO:1 may also contain conservative substitutions of 1, 2, 3, 4, 5, 6, or 7 amino acids in SEQ ID NO:1 as described above.

[0120] In some implementations, the mitochondrial division inhibitory peptide has the structure of formula (Ⅳ'):

[0121] X1-aa1-aa2-aa3-aa4-aa5-aa6-aa7-X2 (IV')

[0122] X1 and X2 may exist simultaneously or not simultaneously. X1 is selected from [G / A / S]-[R / K], R and K, and X2 is selected from [G / A / S]-[I / L / V], G, A and S. aa1-aa7 are defined as in equation (Ⅳ).

[0123] In some embodiments of formula (Ⅳ'), the mitochondrial mitochondrial mitochondrial inhibitory peptide is X1-[D / E]-[L / I / M / V / F]-[L / I / M / V / F]-P-[R / K]-[G / A / T / S]-[T / S / A / G]-X2. In other embodiments, the mitochondrial mitochondrial inhibitory peptide is X1-DLLPRG-[T / S / A / G]-X2. In still other embodiments, the mitochondrial mitochondrial inhibitory peptide is X1-DLLPRG-[S / T]-X2. In some embodiments, X1 and X2 are present simultaneously, where X1 is selected from [G / A / S]-[R / K], R, and K, and X2 is selected from [G / A / S]-[I / L / V], G, A, and S. In some embodiments, X1 is selected from [G / A / S]-[R / K], R, and K, and X2 is absent. In some implementations, X1 is not present, and X2 is selected from [G / A / S]-[I / L / V], G, A, and S.

[0124] Membrane-penetrating peptides

[0125] In some embodiments, the cell-penetrating peptide is fused with a mitochondrial fission inhibitor peptide as described above. Without being bound by any theory, the mitochondrial fission inhibitor peptide fused with the cell-penetrating peptide can cross the blood-brain barrier and localize to the intracellular and / or organelle (e.g., mitochondria). As used herein, a “cell-penetrating peptide (CPP)” refers to a class of peptides capable of penetrating phospholipid membranes (e.g., phospholipid bilayers, lipid microclusters, organelle membranes, vesicle membranes, and cell membranes), typically acting as a delivery carrier linked to a substance of interest (e.g., a mitochondrial fission inhibitor peptide as described above) to deliver the substance of interest to a desired site, such as within the cell or organelle. Preferably, the CPP can cross epithelial cells and the blood-brain barrier. CPPs that can be used include, but are not limited to, cationic CPPs (e.g., those rich in basic amino acids, such as arginine (R) and lysine (K)), amphiphilic CPPs, and hydrophobic CPPs. CPPs known in the art include, but are not limited to, amino acids 47-57 of the transactivator protein (TAT) of human immunodeficiency virus HIV-1. 47-57 (SEQ ID NO:5) and amino acids TAT ​​at positions 49-57 49-57(SEQ ID NO:6), Transportan (SEQ ID NO:11), penetratin derived from Drosophila antennal foot protein (SEQ ID NO:13), polyarginine (e.g., SEQ ID NO:14), polylysine (e.g., containing 3-50 consecutive or non-consecutive lysine residues), etc. Exemplary transmembrane peptides may comprise amino acid sequences selected from SEQ ID NO:3-SEQ ID NO:14, as well as other peptides containing polyarginine, said other polyarginine peptides containing 3-50 consecutive or non-consecutive arginine residues.

[0126] In some embodiments, the membrane-penetrating peptide comprises an amino acid sequence selected from SEQ ID NO:3-SEQ ID NO:14, as well as other peptides containing polyarginine, said other polyarginine-containing peptides comprising 3-50 consecutive or discontinuous arginine residues. In a preferred embodiment, the membrane-penetrating peptide comprises an amino acid sequence selected from SEQ ID NO:3-SEQ ID NO:9. In a specific embodiment, the membrane-penetrating peptide comprises an amino acid sequence of SEQ ID NO:3 or SEQ ID NO:5.

[0127] The membrane-penetrating peptide can be located at the N-terminus and / or C-terminus of the mitochondrial division inhibitory peptide. In some embodiments, in formula (III), p is 1, q is 0, and the membrane-penetrating peptide is located at the N-terminus of the mitochondrial division inhibitory peptide. In other embodiments, in formula (III), p is 0, q is 1, and the membrane-penetrating peptide is located at the C-terminus of the mitochondrial division inhibitory peptide. In still other embodiments, in formula (III), p is 1, q is 1, and the membrane-penetrating peptide is located at both the N-terminus and C-terminus of the mitochondrial division inhibitory peptide.

[0128] connector

[0129] In some embodiments, the membrane-penetrating peptide and the mitochondrial mitochondrial repression peptide are linked via a linker. In some embodiments, the linker is a portion containing an active group, which is linked to both the membrane-penetrating peptide and the mitochondrial mitochondrial repression peptide, respectively. In a preferred embodiment, the linker is an amino acid or peptide linker. Preferred linkers contain 1-20 amino acids, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 amino acids. Exemplary linkers may contain glycine (G), alanine (A), serine (S), and combinations thereof. For example, the linker may contain an amino acid sequence selected from the following: (G) n G n A, (GS) n G n S m 、(G n S) m Gn S, where n and m are each independently selected from integers from 1 to 20.

[0130] In some embodiments, the linker comprises 1-20 amino acids. In some embodiments, the linker is selected from (G). n G n A, (GS) n G n S m 、(G n S) m G n S, where n and m are each independently selected from integers from 1 to 20. In some preferred embodiments, the connector is selected from G, GG, GGG, GGGG, GGSG, GGGGS, GGGSG, and GSSSG. In one specific embodiment, the connector is GG.

[0131] Specific embodiments of the compound of formula (Ⅲ)

[0132] In some implementations, p is 1, q is 0, and Z is a mitochondrial division inhibitory peptide; the compound of formula (III) has the structure of formula (III-1):

[0133] CP1-Linker1-Z (Ⅲ-1)

[0134] CP1 is a transmembrane peptide, and Linker1 is either absent or a linker.

[0135] In some implementations, p is 0, q is 1, and Z is a mitochondrial division inhibitory peptide; the compound of formula (III) has the structure of formula (III-1):

[0136] Z-Linker2-CP2 (Ⅲ-2)

[0137] CP2 is a membrane-penetrating peptide, while Linker2 is either absent or a linker.

[0138] In some implementations, p is 1, q is 1, and Z is a mitochondrial division inhibitory peptide; the compound of formula (III) has the structure of formula (III-3):

[0139] CP1-Linker1-Z-Linker2-CP2 (III-3)

[0140] CP1 and CP2 are each independently transmembrane peptides; Linker1 and Linker2 are either not present or are linkers.

[0141] In some implementations, p and q are both 0, Z is a mitochondrial division inhibitory peptide, and the compound of formula (III) is Z.

[0142] In some embodiments, Z has the structure of formula (Ⅳ). In some embodiments, Z is [D / E]-[L / I / M / V / F]-[L / I / M / V / F]-P-[R / K]-[G / A / T / S]-[T / S / A / G]. In some embodiments, Z is DLLPRG-[T / S / A / G]. In a specific embodiment, Z has the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2.

[0143] In some embodiments, Z has the structure of formula (Ⅳ'). In other embodiments, Z is X1-[D / E]-[L / I / M / V / F]-[L / I / M / V / F]-P-[R / K]-[G / A / T / S]-[T / S / A / G]-X2. In some embodiments, Z is X1-DLLPRG-[T / S / A / G]-X2. In some embodiments, Z is X1-DLLPRG-[S / T]-X2. In some embodiments, X1 and X2 are present simultaneously, where X1 is selected from [G / A / S]-[R / K], R, and K, and X2 is selected from [G / A / S]-[I / L / V], G, A, and S. In some embodiments, X1 is selected from [G / A / S]-[R / K], R, and K, and X2 is not present. In some implementations, X1 is not present, and X2 is selected from [G / A / S]-[I / L / V], G, A, and S.

[0144] In some embodiments, the membrane-penetrating peptide comprises an amino acid sequence selected from SEQ ID NO:3-SEQ ID NO:14 or other amino acid sequences, said other amino acid sequences comprising 3-50 consecutive or discontinuous arginine residues. In one specific embodiment, the membrane-penetrating peptide comprises an amino acid sequence selected from SEQ ID NO:3-SEQ ID NO:6. In a preferred embodiment, the membrane-penetrating peptide comprises an amino acid sequence of SEQ ID NO:3 or SEQ ID NO:5.

[0145] In some embodiments, the linker comprises 1-20 amino acids. In some embodiments, the linker is selected from (G). n G n A, (GS) n G n S m 、(G n S) m G n S, where n and m are each independently selected from integers from 1 to 20. In some preferred embodiments, the connector is selected from G, GG, GGG, GGGG, GGSG, GGGGS, GGGSG, and GSSSG. In one specific embodiment, the connector is GG.

[0146] In one specific embodiment of formula (Ⅲ-1), CP1 comprises the amino acid sequence of SEQ ID NO:3 (RRRQRRKKRGY) or SEQ ID NO:5 (YGRKKRRQRRR), Linker1 is GG, Z has the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2, and the compound of formula (Ⅲ) has an amino acid sequence selected from the following: RRRQRRKKRGYGGDLLPRGS (SEQ ID NO:15), RRRQRRKKRGYGGDLLPRGT (SEQ ID NO:16), YGRKKRRQRRRGGDLLPRGS (SEQ ID NO:17), and YGRKKRRQRRRGGDLLPRGT (SEQ ID NO:18). In one specific embodiment, the compound of formula (Ⅲ) is P110, which has the amino acid sequence of SEQ ID NO:15 or SEQ ID NO:17.

[0147] In other embodiments, both p and q are 0, and the compound of formula (III) is [D / E]-[L / I / M / V / F]-[L / I / M / V / F]-P-[R / K]-[G / A / T / S]-[T / S / A / G]. In some embodiments, the compound of formula (III) is DLLPRG-[T / S / A / G]. In one specific embodiment, the compound of formula (III) has the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2.

[0148] In some embodiments, the compound of formula (III) comprises modifications at the amino terminus and / or carboxyl terminus. In one embodiment, the modification at the amino terminus is acetylation of the terminal amino group. In one embodiment, the modification at the carboxyl terminus is amidation of the terminal carboxyl group. In one embodiment, the compound of formula (III) is P110, having the amino acid sequence of SEQ ID NO:15 or SEQ ID NO:17, and comprising amidation modification of the terminal carboxyl group (i.e., the carboxyl group on the terminal serine residue).

[0149] Preparation method of compound of formula (III)

[0150] A description of compounds of formula (III) can be found, for example, in WO 2012 / 158624, the entire contents of which are incorporated herein by reference. Those skilled in the art will understand that compounds of formula (III) can be, for example, polypeptides or peptide conjugates. Methods for preparing polypeptides and peptide conjugates are well known to those skilled in the art. For example, methods for preparing compounds of formula (III) may include, but are not limited to: obtaining a nucleic acid encoding a compound of formula (III) using recombinant DNA technology and expressing said nucleic acid in a suitable system (e.g., a host cell); chemical synthesis; and covalently conjugating a mitochondrial division inhibitory peptide to a transmembrane peptide using a linker containing an active group. In some embodiments, P110 is commercially available.

[0151] 2. Opioid dependence

[0152] As used in this article, "opioid dependence" refers to a condition resulting from long-term use of opioid analgesics. Opioid dependence can manifest as physical and psychological dependence. Typically, its symptoms may include increased tolerance to opioid analgesics, as well as withdrawal symptoms (e.g., physical and psychological withdrawal symptoms) after stopping or reducing the use of opioid analgesics.

[0153] As used herein, "tolerance" to opioid analgesics refers to the need to increase the dose of the opioid analgesic to achieve the same analgesic effect. Generally, subjects using opioid analgesics long-term (e.g., patients with chronic pain) are prone to developing tolerance, leading to a decrease in the analgesic effect of opioid analgesics over time. In some implementations, Mdivi-1 reduces tolerance to morphine caused by long-term use, thereby enhancing the analgesic effect of morphine.

[0154] In this article, “withdrawal” from the use of opioid analgesics refers to stopping or reducing the use of opioid analgesics after long-term use.

[0155] As used in this article, “withdrawal symptoms” regarding the use of opioid analgesics can include physical and psychological discomfort symptoms that occur after withdrawal from opioid analgesics.

[0156] Acute withdrawal symptoms are generally considered to occur during early withdrawal after cessation or significant reduction of opioid analgesic use (e.g., hours or days after the last dose). Acute withdrawal symptoms can include, but are not limited to, sweating, tremors, fever, chills, fatigue, nausea, vomiting, diarrhea, dilated pupils, recurrent chills, runny nose and tearing, muscle pain or cramps, gastrointestinal spasms, tachycardia, mood disturbances, anxiety, depression, and sleep disturbances. In some implementations, acute withdrawal symptoms are induced with opioid receptor antagonists (e.g., naloxone).

[0157] Even days, months, or years after the acute withdrawal symptoms subside, a range of physical and psychological symptoms may persist, known as "delayed withdrawal symptoms." Delayed withdrawal symptoms can include, for example, sleep disturbances, pain, mood disturbances, gastrointestinal symptoms, cravings, general weakness, anxiety, social avoidance, and depression. Generally, both acute and delayed withdrawal symptoms can include both physical and psychological symptoms. It is generally believed that acute withdrawal symptoms are predominantly physical, while delayed withdrawal symptoms are predominantly psychological. Physical symptoms can include, for example, sweating, insomnia, muscle pain, runny nose and tearing, tremors, fever, chills, diarrhea, dilated pupils, weakness, and sleep disturbances. Psychological symptoms can include, for example, cravings, anxiety, depression, and social avoidance.

[0158] In this article, "prevention or treatment of opioid dependence" can include preventing, reducing, alleviating, or eliminating opioid dependence symptoms or slowing the progression of related symptoms. For example, the efficacy of "prevention or treatment of opioid dependence" can manifest as the prevention, reduction, alleviation, or elimination of opioid withdrawal symptoms (e.g., acute and prolonged withdrawal symptoms) or the reduction, inhibition, or elimination of tolerance to opioid analgesics.

[0159] In some embodiments, the drug is used to inhibit mitochondrial fragmentation of dopaminergic neurons in the ventral tegmentum of the midbrain caused by long-term use of opioid analgesics. In some embodiments, the drug is used to protect dopaminergic neurons in the ventral tegmentum of the midbrain from neuronal damage caused by long-term use of opioid analgesics. In some embodiments, the drug is used to prevent or reduce dependence on opioid analgesics in a subject. In some embodiments, the drug is used to prevent or reduce withdrawal symptoms induced by opioid analgesic withdrawal. In one specific embodiment, the mitochondrial fragmentation inhibitor is Mdivi-1, and the opioid analgesic is morphine. In one specific embodiment, Mdivi-1 reduces somatic symptoms induced by naloxone-induced acute morphine withdrawal. In one specific embodiment, Mdivi-1 reduces prolonged withdrawal symptoms induced by spontaneous morphine withdrawal, such as reducing anxiety levels, depression levels, and social avoidance levels.

[0160] In some embodiments, the drug is used to reduce opioid tolerance caused by long-term use of opioid analgesics. In some embodiments, the drug is used to enhance the analgesic effect of opioid analgesics in subjects using opioid analgesics long-term.

[0161] In some embodiments, the mitochondrial division inhibitor is selected from compounds of formula (I) and their pharmaceutically acceptable salts as described above, compounds of formula (III) and their pharmaceutically acceptable salts, and combinations thereof.

[0162] In some other embodiments, the mitochondrial division inhibitor is a compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof. In one embodiment, the mitochondrial division inhibitor is Mdivi-1 or a pharmaceutically acceptable salt thereof. In a specific embodiment, the mitochondrial division inhibitor is Mdivi-1.

[0163] In other embodiments, the mitochondrial splitting inhibitor is a compound of formula (III) as described above or a pharmaceutically acceptable salt thereof. In one embodiment, the mitochondrial splitting inhibitor has the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2. In yet another embodiment, the mitochondrial splitting inhibitor has the amino acid sequence of SEQ ID NO:15-SEQ ID NO:18. In a specific embodiment, the mitochondrial splitting inhibitor is P110, having the amino acid sequence of SEQ ID NO:15 or SEQ ID NO:17, and optionally comprising an amidation modification of the terminal carboxyl group.

[0164] In some embodiments, the opioid analgesic is selected from morphine, dihydromorphine, diacetylmorphine, hydromorphone, hydroxymorphone, oxycodone, hydrocodone, codeine, dihydrocodeine, buprenorphine, methadone, fentanyl, remifentanil, sufentanil, buprenorphine, meperidine, dezocine, butorphanol, bremazocine, carfentanyl, cyclazocine, enadoline, eseroline, ethylmorphine, etonitazine, etorphanol, levofenphene, levofenol, nalbuphine, nicomorphine, pentazocine, phenazocine, picenadol, tramadol, tapentadol, their pharmaceutically acceptable salts, and combinations thereof. In some embodiments, the opioid analgesic is selected from morphine, buprenorphine, fentanyl, oxycodone, methadone, their pharmaceutically acceptable salts, and combinations thereof. In some embodiments, the opioid analgesic is morphine. In one specific implementation, the opioid analgesic is morphine, and the mitochondrial division inhibitor is Mdivi-1 or P110.

[0165] In some embodiments, the opioid analgesic is used to treat chronic pain, acute pain, postoperative pain, intermittent pain, penetrating pain, tearing pain, mild pain, moderate pain, severe pain, excruciating pain, bone pain, joint pain, soft tissue pain, neuralgia, cancer pain, inflammatory pain, and traumatic pain. In a preferred embodiment, the opioid analgesic is used to treat chronic pain, cancer pain, and inflammatory pain. In some embodiments, the opioid analgesic is used to treat moderate or severe pain, such as moderate or severe pain as defined in the WHO guidelines for the pharmacological and radiotherapeutic management of cancer pain in adults and adolescents (Geneva: World Health Organization; 2018).

[0166] In a further aspect, the present invention also relates to methods for preventing or treating opioid analgesic dependence or enhancing the analgesic effect of opioid analgesics, comprising administering to a subject a mitochondrial splitting inhibitor as described above. In some embodiments, the opioid analgesic is as described above. In some embodiments, the subject is a mammal, such as a human or non-human primate, cattle, pig, horse, sheep, cat, dog, rat, and mouse, particularly a human. In a preferred embodiment, the subject is a person who requires long-term use of opioid analgesics, particularly a patient with cancer, inflammation, or other serious illnesses.

[0167] In some embodiments, the method includes administering a mitochondrial splitting inhibitor during the administration of an opioid analgesic. For example, administering one or more doses of a mitochondrial splitting inhibitor between administrations of two or more doses of an opioid analgesic. In a preferred embodiment, the method includes administering the mitochondrial splitting inhibitor immediately before each dose of the opioid analgesic, for example, 0.5, 1, 2, 3, or 4 hours before administration of the opioid analgesic.

[0168] II. Analgesia

[0169] In another general aspect, the present invention provides a combination comprising:

[0170] (1) One or more mitochondrial division inhibitors as described above; and

[0171] (2) Opioid analgesics as described above.

[0172] In some implementations, the combination is used to provide analgesia to the subject.

[0173] There are no particular limitations on the types of pain that the combination can treat. In some embodiments, the pain is selected from chronic pain, acute pain, postoperative pain, intermittent pain, penetrating pain, tearing pain, mild pain, moderate pain, severe pain, excruciating pain, bone pain, joint pain, soft tissue pain, neuralgia, cancer pain, inflammatory pain, and traumatic pain. In a preferred embodiment, the pain is selected from chronic pain, cancer pain, and inflammatory pain. In some embodiments, the pain is moderate or severe pain. In some embodiments, the pain is moderate or severe pain as defined in the WHO guidelines for the pharmacoscopic and radiotherapeutic management of cancer pain in adults and adolescents. Geneva: World Health Organization; 2018.

[0174] In some embodiments, the mitochondrial division inhibitor is selected from compounds of formula (I) as described above and their pharmaceutically acceptable salts, compounds of formula (III) and their pharmaceutically acceptable salts, and combinations thereof.

[0175] In some embodiments, the mitochondrial division inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, the mitochondrial division inhibitor is Mdivi-1 or a pharmaceutically acceptable salt thereof. In a specific embodiment, the mitochondrial division inhibitor is Mdivi-1.

[0176] In other embodiments, the mitochondrial splitting inhibitor is a compound of formula (III) or a pharmaceutically acceptable salt thereof. In one embodiment, the mitochondrial splitting inhibitor has the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2. In yet another embodiment, the mitochondrial splitting inhibitor has an amino acid sequence selected from SEQ ID NO:15-SEQ ID NO:18. In a specific embodiment, the mitochondrial splitting inhibitor is P110, having the amino acid sequence of SEQ ID NO:15 or SEQ ID NO:17, and optionally comprising an amidation modification of the terminal carboxyl group.

[0177] Those skilled in the art can select appropriate opioid analgesics based on needs (e.g., the patient's physical condition, the type and severity of pain). For cancer pain patients, guidelines such as those developed by the WHO for the pharmacological and radiotherapeutic management of cancer pain in adults and adolescents (Geneva: World Health Organization; 2018) can be consulted to select appropriate opioid analgesics according to the severity of pain. In some embodiments, the opioid analgesic is selected from morphine, dihydromorphine, diacetylmorphine, hydromorphone, hydroxymorphone, oxycodone, hydrocodone, codeine, dihydrocodeine, buprenorphine, methadone, fentanyl, remifentanil, sufentanil, buprenorphine, meperidine, dezocine, butorphanol, bremazocine, carfentanyl, cyclazocine, enadoline, eseroline, ethylmorphine, etonitazine, etorphanol, levofenphene, levofenol, nalbuphine, nicomorphine, pentazocine, phenazocine, picenadol, tramadol, tapentadol, their pharmaceutically acceptable salts, and combinations thereof. In some embodiments, the opioid analgesic is selected from morphine, fentanyl, sufentanil, remifentanil, codeine, dihydrocodeine, oxycodone, methadone, their pharmaceutically acceptable salts, and combinations thereof. In one specific implementation plan, the opioid analgesic is morphine.

[0178] In one specific implementation, the opioid analgesic is morphine, and the mitochondrial division inhibitor is Mdivi-1 or P110.

[0179] In some embodiments, mitochondrial splitting inhibitors and opioid analgesics are present in separate compositions. In one exemplary embodiment, the combination comprises: (1) a first composition comprising one or more mitochondrial splitting inhibitors as described above and optionally a pharmaceutically acceptable carrier; and (2) a second composition comprising an opioid analgesic and optionally a pharmaceutically acceptable carrier. Such combinations may be presented as kits in which the first and second compositions are present in the same or separate portions. In some embodiments, the components of the combination are present in the same composition. Therefore, the present invention also provides a composition comprising a mitochondrial splitting inhibitor and an opioid analgesic, and optionally a pharmaceutically acceptable carrier. In one specific embodiment, the opioid analgesic is morphine, and the mitochondrial splitting inhibitor is Mdivi-1 or P110.

[0180] In some embodiments, the combination may optionally further comprise: (3) a non-opioid analgesic and / or (4) an adjunctive analgesic. Non-opioid analgesics may include, for example, nonsteroidal anti-inflammatory drugs (NSAIDs). Examples of NSAIDs may include, but are not limited to, aspirin, indomethacin, acetaminophen, ibuprofen, phenylbutazone, naproxen, naproxone, diclofenac, nimesulide, rofecoxib, celecoxib, ketoprofen, flurbiprofen, loxoprofen, zaltoprofen, etc. Examples of adjuvant analgesics may include, for example, corticosteroids (e.g., dexamethasone, prednisone), anticonvulsants (e.g., carbamazepine, phenytoin sodium, gabapentin, pregabalin), antidepressants (e.g., amitriptyline, venlafaxine, mesyrel, duloxetine, doxepin, prozac), NMDA receptor antagonists (e.g., dextromethorphan), and hydroxyzine antihistamines. In some embodiments, the non-opioid analgesic is a nonsteroidal anti-inflammatory drug (NSAID). Preferably, the NSAID is selected from aspirin, indomethacin, acetaminophen, ibuprofen, phenylbutazone, naproxen, naproxen, diclofenac, nimesulide, rofecoxib, celecoxib, ketoibuprofen, flurbiprofen, loxoprofen, zaltoprofen, and combinations thereof. In some embodiments, the adjunctive analgesic is selected from corticosteroids, antidepressants, NMDA receptor antagonists, hydroxyzine antihistamines, and combinations thereof. In view of this disclosure, those skilled in the art can select specific non-opioid analgesics and / or adjunctive analgesics based on actual needs (e.g., the subject's physical condition, medical history, type and severity of pain, the specific opioid analgesic chosen, etc.).

[0181] In another aspect, the present invention also provides the use of the said combination in the preparation of a medicament for providing analgesia to a subject. In some embodiments, the medicament is used to treat chronic pain, acute pain, postoperative pain, intermittent pain, penetrating pain, tearing pain, mild pain, moderate pain, severe pain, excruciating pain, bone pain, joint pain, soft tissue pain, neuralgia, cancer pain, inflammatory pain, and traumatic pain; preferably, the medicament is used to treat cancer pain, chronic pain, and inflammatory pain; more preferably, the medicament is used to treat moderate or severe pain.

[0182] In a further aspect, the present invention also provides a method for providing analgesia to a subject, the method comprising administering the above-described combination to the subject. In one specific embodiment, the opioid analgesic is morphine, and the mitochondrial splitting inhibitor is Mdivi-1 or P110.

[0183] In some embodiments, the method includes administering a mitochondrial splitting inhibitor during the administration of an opioid analgesic. For example, administering one or more doses of a mitochondrial splitting inhibitor between administrations of two or more doses of an opioid analgesic. In a preferred embodiment, the method includes administering the mitochondrial splitting inhibitor immediately before each dose of the opioid analgesic, for example, 0.5, 1, 2, 3, or 4 hours before administration of the opioid analgesic.

[0184] In some embodiments, the combination comprising a mitochondrial splitting inhibitor and an opioid analgesic is administered in the same composition. In some embodiments, the combination comprising a mitochondrial splitting inhibitor and an opioid analgesic is administered in two or more compositions.

[0185] In some embodiments, the subject is a mammal. Examples include humans and non-human primates, cattle, pigs, horses, sheep, cats, dogs, rats, and mice, particularly humans. In a preferred embodiment, the subject is a person who requires long-term use of opioid analgesics, particularly a patient with cancer, inflammation, or other serious illnesses.

[0186] Dosage

[0187] The active substance (e.g., mitochondrial fission inhibitors and opioid analgesics as described above) can be administered alone. Alternatively, the active substance can be prepared into a pharmaceutical composition for administration according to methods known in the art. The active substance or pharmaceutical composition can be prepared into dosage forms commonly used in the art, such as powders, tablets, capsules, microspheres (e.g., nanospheres), liposomes, suppositories, patches, and injections. For compounds of formula (III) or their pharmaceutically acceptable salts, a system capable of drug delivery across the blood-brain barrier is preferred, and the system is formulated into a suitable dosage form. Delivery systems capable of crossing the blood-brain barrier include, but are not limited to, nanoparticles (e.g., nanospheres and nanocapsules), liposomes (e.g., multilayer vesicles, large monolayer vesicles, and small monolayer vesicles), and niosomes. A description of such delivery systems can be found, for example, in Pardridge WM., FrontAging Neurosci. (2020) 11:373.

[0188] The mitochondrial division inhibitors described herein can be prepared into dosage forms suitable for administration using methods known to those skilled in the art. Suitable dosage forms may be, for example, oral dosage forms (e.g., oral tablets and oral solutions) or injectable dosage forms. In some embodiments, the medicine for preventing or treating opioid analgesic dependence or enhancing the analgesic effect of opioid analgesics is an oral dosage form or injectable dosage form. In some embodiments, the medicine for providing analgesia to a subject is an oral dosage form or injectable dosage form.

[0189] In embodiments involving the combination of the present invention, mitochondrial splitting inhibitors and opioid analgesics can be administered in the same or different dosage forms. For example, opioid analgesics can be formulated as oral dosage forms (e.g., controlled-release tablets, extended-release tablets, oral solutions), injections, or patches, such as morphine sulfate controlled-release tablets, oxycodone hydrochloride controlled-release tablets, oxycodone hydrochloride injection, or fentanyl transdermal patches. Mitochondrial inhibitors can be formulated as oral dosage forms or injections. In one specific embodiment, the opioid analgesic is morphine, and the mitochondrial splitting inhibitor is Mdivi-1 or P110.

[0190] The mitochondrial fission inhibitors, opioid analgesics, or combinations thereof described herein may be administered at the same dose or at different doses. For example, the mitochondrial fission inhibitors, opioid analgesics, or combinations thereof may be administered at one or more doses (e.g., 1, 2, 3, 4, 5, 6 or more doses per day, or 1, 2, 3, 4, 5, 6, 7 or more doses per week).

[0191] The active substance or pharmaceutical composition comprising it may be administered independently via one or more routes / methods known in the art. Those skilled in the art will understand that the route and / or method of administration varies depending on the desired outcome.

[0192] Generally, routes of administration can include, but are not limited to, parenteral and non-parenteral administration. "Parenteral administration" refers to administration methods other than enteric and local administration, typically via injection, including but not limited to intravenous, intramuscular, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions. "Non-parenteral administration" can be, for example, local, epidermal, or mucosal routes, such as intranasal, oral, vaginal, rectal, sublingual, or local administration. Preferred routes of administration include intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal, or other parenteral routes.

[0193] The combinations of the present invention can be administered to a subject using methods known in the art. Exemplary administration methods may include, but are not limited to, injection, inhalation, oral, transdermal, chewing, and sublingual administration. In embodiments for providing analgesia, patient-controlled analgesia (PCA) administration may also be selected, such as subcutaneous PCA, intravenous PCA, epidural PCA, or regional nerve block PCA.

[0194] Exemplary embodiments of the present invention include:

[0195] 1. Use of mitochondrial fission inhibitors in the preparation of medicaments for the prevention or treatment of opioid dependence or for enhancing the analgesic effect of opioid analgesics.

[0196] 2. According to the use described in paragraph 1, the mitochondrial division inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof, the compound of formula (I) having the following structure:

[0197]

[0198] Where R 1 Selected from: hydrogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 cycloalkyl, C 1-6 Heteroalkyl, C 2-6 Heterene, C 2-6 heteroyne group and C 3-8 Heterocyclic alkyl groups;

[0199] R 2 R 3 R 4 and R 5 Each is independently selected from: hydrogen, C 1-6 Alkyl, C 1-6 Heteroalkyl groups and halogens;

[0200] The alkyl, alkenyl, ynyl, cycloalkyl, heteroalkyl, heteroalkenyl, heteroynyl, heterocyclic, and alkyl groups are optionally substituted with one or more of the following groups: halogen, C 1-6 Alkyl, cyano, C 1-6 Alkoxy, C 1-6 Alkylthio, hydroxyl, SH, SO4, C 3-8 cycloalkyl, C 3-8 Heterocyclic alkyl, amino, amide, oxo (O=) and nitro.

[0201] 3. According to the use described in item 1 or 2, the compound of formula (I) has the structure of formula (II):

[0202]

[0203] 4. According to the use described in paragraph 1, the mitochondrial division inhibitor is a compound of formula (III) or a pharmaceutically acceptable salt thereof, the compound of formula (III) having the following structure:

[0204] (CP1-Linker1) p -Z-(Linker2-CP2) q (III)

[0205] in

[0206] CP1 and CP2 are each independently transmembrane peptides;

[0207] Linker1 and Linker2 either do not exist independently, or they are connectors;

[0208] Z is a mitochondrial division inhibitory peptide;

[0209] p and q are each independently 0 or 1; preferably, p is 0 and q is 0 or 1; or p is 1 and q is 0.

[0210] 5. According to the use in item 4, the mitochondrial division inhibitory peptide is [D / E]-[L / I / M / V / F]-[L / I / M / V / F]-P-[R / K]-[G / A / T / S]-[T / S / A / G].

[0211] 6. According to the use of item 5, wherein the mitochondrial division inhibitory peptide is DLLPRG-[T / S / A / G]; preferably, the mitochondrial division inhibitory peptide is DLLPRG-[S / T].

[0212] 7. Depending on the purpose of item 5 or 6, where p is 1, q is 0, and CP1 and Linker1 are as defined in item 4; or

[0213] p is 0, q is 1, and CP2 and Linker2 are defined as in item 4.

[0214] 8. According to the use of item 4, wherein the mitochondrial division inhibitory peptide is X1-[D / E]-[L / I / M / V / F]-[L / I / M / V / F]-P-[R / K]-[G / A / T / S]-[T / S / A / G]-X2, wherein X1 and X2 are present simultaneously or not simultaneously, X1 is selected from [G / A / S]-[R / K], R and K, and X2 is selected from [G / A / S]-[I / L / V], G, A and S; preferably, the mitochondrial division inhibitory peptide is X1-DLLPRG-[T / S / A / G]-X2; more preferably, the mitochondrial division inhibitory peptide is X1-DLLPRG-[S / T]-X2.

[0215] 9. Based on the purpose of any one of items 4-8, among which

[0216] The membrane-penetrating peptide comprises an amino acid sequence selected from SEQ ID NO:3-SEQ ID NO:14 or other amino acid sequences, wherein the other amino acid sequences comprise 3-50 consecutive or non-consecutive arginine residues; preferably, the membrane-penetrating peptide comprises an amino acid sequence selected from SEQ ID NO:3-SEQ ID NO:6;

[0217] The linker contains 1-20 amino acids; preferably, the linker is selected from (G). n G n A, (GS) n G n S m 、(G n S) m G n S, where n and m are each independently selected from integers from 1 to 20; more preferably, the connector is selected from G, GG, GGG, GGGG, GGSG, GGGGS, GGGSG and GSSSG.

[0218] 10. According to the use of item 7, wherein the compound of formula (III) has an amino acid sequence selected from SEQ ID NO:15-SEQ ID NO:18; preferably, the compound of formula (III) has the amino acid sequence of SEQ ID NO:15.

[0219] 11. Depending on the use of item 5 or 6, where both p and q are 0.

[0220] 12. According to the use of item 11, the compound of formula (III) has the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2.

[0221] 13. According to the purpose of any one of items 1-12, among which

[0222] The drug is used to inhibit the fragmentation of mitochondria in dopaminergic neurons in the ventral tegmental area of ​​the midbrain caused by long-term use of opioid analgesics; or

[0223] The drug is used to protect dopaminergic neurons in the ventral tegmental area of ​​the midbrain from neuronal damage caused by long-term use of opioid analgesics; or

[0224] The drug is used to prevent or reduce the subject's dependence on opioid analgesics; or

[0225] The drug is used to reduce opioid tolerance caused by long-term use of opioid analgesics; or

[0226] The drug is used to enhance the analgesic effect of opioid analgesics in subjects who have been using opioid analgesics for a long time.

[0227] 14. An assembly comprising:

[0228] (1) One or more mitochondrial division inhibitors as defined in any of items 1-12; and

[0229] (2) Opioid analgesics;

[0230] Optionally, the combination may further comprise non-opioid analgesics and / or adjunctive analgesics;

[0231] The non-opioid analgesic is a nonsteroidal anti-inflammatory drug (NSAID); preferably, the NSAID is selected from aspirin, indomethacin, acetaminophen, ibuprofen, phenylbutazone, naproxen, naproxen, diclofenac, nimesulide, rofecoxib, celecoxib, ketoibuprofen, flurbiprofen, loxoprofen, zaltoprofen, and combinations thereof;

[0232] The adjunctive analgesic is selected from corticosteroids, antidepressants, NMDA receptor antagonists, hydroxyzine antihistamines, and combinations thereof.

[0233] 15. The combination according to item 14, wherein the mitochondrial division inhibitor and the opioid analgesic are present in the same or different compositions.

[0234] 16. The use according to any one of items 1-13 or a combination of items 14 or 15, wherein said opioid analgesic is selected from morphine, dihydromorphine, diacetylmorphine, hydromorphone, oxymorphone, oxycodone, hydrocodone, codeine, dihydrocodeine, buprenorphine, methadone, fentanyl, remifentanil, sufentanil, buprenorphine, meperidine, dezocine, butorphanol, bremazocine, carfentanyl, cyclazocine, enadoline, eseroline, ethyl... The opioid analgesic is selected from morphine, etonitazine, etorphine, levofenpheniramine, nalbuphine, nicomorphine, pentazocine, phenazocine, picenadol, tramadol, tapentadol, pharmaceutically acceptable salts thereof, and combinations thereof; preferably, the opioid analgesic is selected from morphine, fentanyl, sufentanil, remifentanil, codeine, dihydrocodeine, oxycodone, methadone, pharmaceutically acceptable salts thereof, and combinations thereof; more preferably, the opioid analgesic is morphine.

[0235] 17. Use in the preparation of a medicament for providing analgesia to a subject, according to any combination of items 14-16.

[0236] 18. According to the use of item 17, the drug is used to treat chronic pain, acute pain, postoperative pain, intermittent pain, penetrating pain, tearing pain, mild pain, moderate pain, severe pain, extreme pain, bone pain, joint pain, soft tissue pain, neuralgia, cancer pain, inflammatory pain, and traumatic pain; preferably, the drug is used to treat cancer pain, chronic pain, and inflammatory pain; more preferably, the drug is used to treat moderate or severe pain.

[0237] 19. According to the use of item 17 or 18, the subject is a mammal; preferably, the subject is a person who needs long-term use of opioid analgesics, such as a patient with cancer, inflammation or other serious illness.

[0238] 20. The use of any combination of any of the items 14-16 or any of the items 17-19, wherein the mitochondrial division inhibitor is administered during administration of the opioid analgesic.

[0239] Beneficial effects

[0240] Long-term use of opioid analgesics can cause serious side effects, such as physical and psychological dependence, and can also affect the analgesic effect. Existing methods for addressing these problems include, for example, using neurotransmitter receptor antagonists (e.g., dopamine receptor antagonists (see, for example, WO 2020 / 055725) and NMDA receptor antagonists (see, for example, WO 2020 / 106454) to control neuronal function and thus improve analgesia, or developing novel non-addictive analgesics to avoid drug dependence. However, on the one hand, neurotransmitter receptor antagonists can disrupt the normal physiological function of receptors; on the other hand, antagonists may have off-target effects. Therefore, the use of neurotransmitter receptor antagonists may lead to significant side effects. Furthermore, the analgesic effect of currently developed non-addictive analgesics is often lower than that of opioid analgesics at the same dosage, and most are still in the preclinical research stage.

[0241] The inventors discovered that mitochondrial homeostasis was disrupted in dopaminergic neurons of the ventral tegmental area (VTA) of the midbrain in mice subjected to long-term morphine administration, and that neuronal mitochondria exhibited fragmentation. This invention uses mitochondrial fission inhibitors to prevent or alleviate opioid dependence caused by long-term opioid analgesic use, reduce analgesic tolerance, and alleviate physical and psychological symptoms after opioid withdrawal. Furthermore, mitochondrial fission inhibitors can enhance the analgesic effect of opioid analgesics. Compared to existing technologies, mitochondrial fission inhibitors do not act on neurotransmitter receptors, have fewer side effects, and simultaneously achieve the beneficial effects of reducing drug dependence and enhancing the analgesic effect of opioids.

[0242] Example

[0243] A further understanding of the invention can be obtained by referring to some specific embodiments given herein, which are for illustrative purposes only and are not intended to limit the scope of the invention in any way. Obviously, various modifications and variations can be made to the invention without departing from its spirit; therefore, such modifications and variations are also within the scope of protection claimed in this application.

[0244] Unless otherwise specified, all reagents and instruments used in the examples section are commercially available.

[0245] Experimental Materials, Methods and Statistical Analysis

[0246] 1. Reagents

[0247] Morphine Hydrochloride Injection (Shenyang No. 1 Pharmaceutical Co., Ltd., 10mg / ml, batch number: 161109-1)

[0248] Compound Mdivi-1 (Sigma-Aldrich, lot number: M0199)

[0249] Peptide P110 (Tocris, batch number: Cat. No. 6897; SEQ ID NO: 17, with the carboxyl group of the terminal Ser group being amidated)

[0250] Control peptide TAT (47-57) (Bachem Americas, lot number: 368000; SEQ ID NO:5)

[0251] Naloxone hydrochloride (China National Institutes for Food and Drug Control, batch number: NLX6-T6SY)

[0252] 2. Laboratory animals and grouping

[0253] Clean-grade adult male C57 / BL6 mice (wild-type mice) or mice specifically expressing Cre recombinase for dopamine neurons (TH-Cre) (provided by the Animal Center of the Chinese Academy of Sciences) (TH-Cre mice express Cre recombinase under the drive of the tyrosine hydroxylase (TH) promoter; specific tracing of VTA dopamine neuron mitochondria can be achieved by injecting Cre-dependent mitochondrial fluorescent tracer virus into the ventral tegmental area (VTA) of the midbrain of TH-Cre mice), 8-10 weeks old, 25-28g. Animals were housed in our laboratory animal room for 7 days prior to surgery, with free access to food, a room temperature of 20-22℃, humidity of 55±5%, and a 12-hour day-night cycle. Experiments began one week after the animals acclimatized to the environment. Mice were randomly divided into groups of 12-15.

[0254] 3. Administration method

[0255] Morphine escalation administration: Mice were given escalating intraperitoneal injections of morphine twice daily (at 10:00 AM and 10:00 PM) for 5 consecutive days in their cages: 10 mg / kg on day 1, 20 mg / kg on day 2, 40 mg / kg on day 3, 60 mg / kg on day 4, and 60 mg / kg on day 5.

[0256] Morphine was administered via subcutaneous injection into the back: 10 mg / kg, once daily for 6 consecutive days.

[0257] Control group (Vehicle) mice were administered an equal volume of corn oil via intraperitoneal injection.

[0258] Mdivi-1 group mice were administered Mdivi-1 (dissolved in corn oil) via intraperitoneal injection.

[0259] 4. Data processing and statistical analysis

[0260] Data were analyzed and processed using SPSS 16.0 statistical software, expressed as mean ± standard error. The values ​​are indicated by ) . Differences between groups were compared using t-tests or rank-sum tests; statistical tests were performed using two-way repeated measures ANOVA, with pairwise comparisons using the Bonferroni method; the cumulative probability distribution analysis of mitochondrial area was conducted using the KS-test. P < 0.05 was considered significant.

[0261] Example 1: Mdivi-1 and P110 inhibit mitochondrial fragmentation in dopaminergic neurons in the ventral tegmental area of ​​the midbrain after long-term morphine use.

[0262] 1.1 Long-term morphine use leads to increased mitochondrial fragmentation in dopaminergic neurons in the ventral tegmental area of ​​the midbrain.

[0263] Cre-dependent mitochondrial fluorescently traced virus was injected into the ventral tegmental area (VTA) of the midbrain of TH-Cre mice to achieve specific tracing of mitochondria in VTA dopamine neurons. Two weeks after viral expression, mice were divided into a morphine group (Mor) and a control group (Sal). The morphine group received escalating doses of morphine, while the control group received an equal volume of saline. Nine hours after the last escalating dose injection, mouse brain tissue was harvested, fixed, and sliced. Dendrites of VTA neurons were imaged using a laser confocal microscope, and the mitochondrial area in the dopaminergic neuronal dendrites was measured. Smaller mitochondrial areas indicated higher fragmentation. Results are shown in... Figure 1A .

[0264] Depend on Figure 1AAs can be seen, compared with the saline control group (Sal), the mean mitochondrial area in the dendrites of dopaminergic neurons in the ventral tegmental region of mice in the morphine group (Mor) was significantly reduced (left, bar chart), and the cumulative distribution frequency was significantly shifted to the left (right, cumulative probability distribution curve); indicating that long-term use of morphine causes mitochondrial fragmentation of dopaminergic neurons in the ventral tegmental region of the midbrain of mice.

[0265] 1.2 Mdivi-1 inhibits mitochondrial fragmentation of dopaminergic neurons in the ventral tegmental area of ​​the midbrain after long-term morphine use.

[0266] Cre-dependent mitochondrial fluorescent tracer virus was injected into the ventral midbrain tegmental area (VTA) of TH-Cre mice to achieve specific mitochondrial tracing of VTA dopamine neurons. Two weeks after viral expression, the mice were divided into the Mdivi-1 group (Mdivi-1) and the control group (Vehicle) and administered morphine in escalating doses in cages. One hour before each morphine injection, the mice were intraperitoneally injected with 50 mg / kg Mdivi-1 (Mdivi-1 group) or an equal volume of corn oil (control group).

[0267] Nine hours after the last incremental dose injection, mouse brain tissue was harvested, fixed, and sliced. Dendritic cells of neurons in the ventral tegmental area (VTA) of the midbrain were imaged using a laser confocal microscope, and the mitochondrial area in the dopaminergic neuron dendrites was measured. A smaller mitochondrial area indicated a higher degree of fragmentation. Results are shown in... Figure 1B .

[0268] Depend on Figure 1B As can be seen, compared with the control group, the mean mitochondrial area in the dendrites of dopaminergic neurons in the ventral tegmental region of mice in the Mdivi-1 group was significantly increased (left, bar chart), and the cumulative distribution frequency was significantly shifted to the right (right, cumulative probability distribution curve); indicating that Mdivi-1 reduces mitochondrial fragmentation of dopaminergic neurons in the ventral tegmental region of the midbrain after long-term morphine use, and that Mdivi-1 can protect dopaminergic neurons and avoid neuronal damage caused by long-term morphine use.

[0269] 1.3 P110 inhibits mitochondrial fragmentation of dopaminergic neurons in the ventral tegmental area of ​​the midbrain after long-term morphine use.

[0270] Cre-dependent mitochondrial fluorescent tracer virus was injected into the ventral midbrain tegmental area (VTA) of TH-Cre mice to achieve specific mitochondrial tracing of VTA dopamine neurons; two weeks after virus expression, the mice were divided into the P110 group (P110) and the control group (TAT). (47-57)In cages, morphine was administered in escalating doses; one hour before each morphine injection, mice were intraperitoneally injected with 1 mg / kg P110 (P110 group) or an equimolar dose of the control peptide TAT. (47-57) Nine hours after the last incremental dose injection, mouse brain tissue was harvested, fixed, and sliced. Dendritic cells of neurons in the ventral tegmental area (VTA) of the midbrain were imaged using a laser confocal microscope, and the mitochondrial area in the dopaminergic neuron dendrites was measured; a smaller mitochondrial area indicated a higher degree of fragmentation. Results are shown in... Figure 1C .

[0271] Depend on Figure 1C As can be seen, compared with the control group, the mean mitochondrial area in the dendrites of dopaminergic neurons in the ventral tegmental region of mice in the P110 group was significantly increased (left, bar chart), and the cumulative distribution frequency was significantly shifted to the right (right, cumulative probability distribution curve); indicating that the Drp1 peptide inhibitor P110 reduces mitochondrial fragmentation of dopaminergic neurons in the ventral tegmental region of the midbrain after long-term morphine use, and that P110 can protect dopaminergic neurons and avoid neuronal damage caused by long-term morphine use.

[0272] Example 2: Mdivi-1 and P110 inhibit analgesia tolerance induced by long-term morphine use and enhance analgesic effect

[0273] Wild-type mice were placed in a hot plate apparatus at a temperature of 50°C. The pain threshold of the mice was measured by observing whether they licked their hind paws or jumped. Mice with a pain threshold of less than 30 seconds were pre-selected for the experiment.

[0274] Mice were randomly divided into Mdivi-1 groups (Mor + Mdivi-1 (12.5 mg / kg), Mor + Mdivi-1 (25 mg / kg), Mor + Mdivi-1 (50 mg / kg), Mor + Mdivi-1 (100 mg / kg)) and a control group (Mor + vehicle). Mice were intraperitoneally injected with 12.5, 25, 50 or 100 mg / kg of Mdivi-1 (Mdivi-1 group) or an equal volume of corn oil (control group). One hour after injection, morphine was subcutaneously injected into the back (10 mg / kg each time, once a day). The pain threshold of the mice was then measured for 6 consecutive days.

[0275] Alternatively, mice can be randomly divided into the P110 group (Mor + P110 (0.2 mg / kg), Mor + P110 (0.5 mg / kg), Mor + P110 (1 mg / kg) and the control group (Mor + TAT). (47-57)Mice were intraperitoneally injected with 0.2, 0.5, or 1 mg / kg P110 (P110 group) or an equimolar dose of control peptide. One hour after the injection, morphine was administered subcutaneously to the back (10 mg / kg each time, once a day). The pain threshold of the mice was then measured for 6 consecutive days.

[0276] If the mouse showed no pain response within 60 seconds, it was immediately removed, and the pain threshold was calculated based on 60 seconds. After recording the pain threshold, the maximum analgesic effect was calculated, which is usually expressed as the percentage of the maximum possible effect (MPE%). The calculation method is: MPE% = (Pain threshold after administration - Pain threshold before administration) / (60 seconds - Pain threshold before administration) * 100%; the higher the MPE%, the better the analgesic effect. The results are shown below. Figure 2A and Figure 2B .

[0277] Depend on Figure 2A It is evident that after prolonged morphine use, the MPE% in both the control and Mdivi-1 groups decreased over time, suggesting that the analgesic effect of morphine decreases with prolonged use, indicating that long-term morphine use induces analgesic tolerance in mice. Compared to the control group, the decreasing trend of MPE% over time was significantly inhibited in mice administered 50 and 100 mg / kg Mdivi-1, indicating that Mdivi-1 can significantly improve analgesic tolerance induced by long-term morphine use, and a clear dose-response relationship exists. Furthermore, on days 2, 3, 4, 5, and 6 after morphine injection, the MPE% in mice administered 50 and 100 mg / kg Mdivi-1 was significantly higher than that in the control group, suggesting that administration of Mdivi-1 can enhance the analgesic effect of morphine during long-term use.

[0278] Depend on Figure 2B It is evident that after prolonged morphine use, the MPE% in both the control and P110 groups decreased over time, suggesting that the analgesic effect of morphine decreases with increasing morphine use duration, indicating that prolonged morphine use induces analgesic tolerance in mice. Compared to the control group, the decreasing trend of MPE% over time was significantly inhibited in mice administered 0.5 and 1 mg / kg P110, indicating that P110 can significantly improve analgesic tolerance induced by prolonged morphine use, and a clear dose-response relationship exists. Furthermore, on days 4, 5, and 6 after morphine injection, the MPE% in mice administered 0.5 and 1 mg / kg P110 was significantly higher than that in the control group, suggesting that administration of P110 can enhance the analgesic effect of morphine during prolonged use.

[0279] Example 3: Mdivi-1 improves physical and psychological symptoms of acute morphine withdrawal.

[0280] 3.1 Mdivi-1 alleviates somatic symptoms of naloxone-induced acute morphine withdrawal.

[0281] To investigate the effect of Mdivi-1 on acute withdrawal symptoms, naloxone was used to induce acute withdrawal symptoms in mice after administering escalating doses of morphine.

[0282] Specifically, wild-type mice were randomly divided into the Mdivi-1 group (Mdivi-1) and the control group (Vehicle). Morphine was administered in escalating doses in cages. One hour before each morphine injection, mice were intraperitoneally injected with 50 mg / kg Mdivi-1 (Mdivi-1 group) or an equal volume of corn oil (control group). Twelve hours after the last morphine injection, mice were intraperitoneally injected with naloxone (1 mg / kg) and then returned to their individual cages. Withdrawal symptoms were recorded via video for 20 minutes. A double-blind statistical analysis was performed offline on somatic symptoms such as wet dog-like tremors, body tremors, backward movements, and piloerection. The results are shown below. Figure 3 .

[0283] Depend on Figure 3 It is evident that, compared with the control group, the number of occurrences of various somatic symptoms (including wet dog-like tremors, body tremors, backward movements, and piloerection) in the Mdivi-1 group mice was significantly reduced, suggesting that naloxone-induced acute morphine withdrawal somatic symptoms were alleviated.

[0284] 3.2 Mdivi-1 alleviates physical discomfort symptoms after spontaneous morphine withdrawal.

[0285] Morphine withdrawal induces physical discomfort symptoms in mice, leading to aversion. We used a conditioned positional aversion (CPA) experiment to study aversion association in mice. The degree of aversion was used to assess the severity of physical discomfort symptoms.

[0286] The Conditioned Place Aversion (CPA) testing system consists of left and right side chambers and a movable partition in the middle. The bottom surfaces of the two side chambers are made of different materials. On the first day, wild-type mice were placed in the Conditioned Place Aversion chamber for 20 minutes and allowed to explore freely (with the partition open). The time spent in the left and right side chambers was recorded as the baseline preference value. Mice with a preference greater than 60% were removed.

[0287] After adaptation training in the testing box, mice were randomly divided into the Mdivi-1 group (Mdivi-1) and the control group (Vehicle). Morphine was administered in escalating doses in cages. One hour before each morphine injection, mice were intraperitoneally injected with 50 mg / kg Mdivi-1 (Mdivi-1 group) or an equal volume of corn oil (control group). Nine hours after each of the last three morphine injections, mice underwent CPA-associated training, learning aversion association for 30 minutes on the restricted side of the training box. On day 6 after training (i.e., 6 days of spontaneous morphine withdrawal), mice were re-exposed to the testing box, allowed free exploration for 20 minutes, and the time (in seconds) spent by each mouse on the restricted and untrained sides was recorded. The CPA score was defined as: time (in seconds) on the restricted side minus time (in seconds) on the untrained side; a higher negative value indicated a stronger degree of aversion. Results are shown below. Figure 4 .

[0288] Depend on Figure 4 It is evident that, compared with the control group, the CPA score of conditioned position aversion after spontaneous morphine withdrawal was significantly reduced in the Mdivi-1 group mice, suggesting that Mdivi-1 can significantly alleviate physical discomfort symptoms after morphine withdrawal.

[0289] 3.3 Mdivi-1 reduces anxiety levels after spontaneous morphine withdrawal.

[0290] Wild-type mice were randomly divided into the Mdivi-1 group (Mdivi-1) and the control group (Vehicle). Morphine was administered in escalating doses in cages. One hour before each morphine injection, mice were intraperitoneally injected with 50 mg / kg Mdivi-1 (Mdivi-1 group) or an equal volume of corn oil (control group). Seven days after the end of the escalating morphine administration (i.e., 7 days of spontaneous morphine withdrawal), the mice were placed in a quiet, dimly lit room in the center of an elevated cross maze, facing open arms, and allowed to explore freely for 6 minutes. The mice's behavior was recorded by a camera mounted above the maze. The total time (in seconds) spent in the open arms was analyzed using an animal behavior tracking and analysis system; shorter time spent in the open arms was associated with higher anxiety levels. Results are shown below. Figure 5 .

[0291] Depend on Figure 5 It is evident that, compared with the control group, the time spent in the elevated cruciate arm of the mice was significantly increased, suggesting that Mdivi-1 can significantly reduce anxiety levels after morphine withdrawal.

[0292] 3.4 Mdivi-1 reduces depressive levels after spontaneous morphine withdrawal.

[0293] Wild-type mice were randomly divided into the Mdivi-1 group (Mdivi-1) and the control group (Vehicle). Morphine was administered in escalating doses in cages. One hour before each morphine injection, mice were intraperitoneally injected with 50 mg / kg Mdivi-1 (Mdivi-1 group) or an equal volume of corn oil (control group). On day 8 after the end of the escalating morphine administration (i.e., 8 days of spontaneous morphine withdrawal), the mice were secured approximately 1 cm from the tail tip with medical tape and suspended upside down about 15 cm from the ground. A 6-minute video recording was simultaneously started for analysis. The animals struggled to overcome the abnormal body position, but after a period of struggle, they exhibited intermittent periods of stillness. The duration of stillness (in seconds) within 6 minutes for each group of mice was analyzed using an animal behavior tracking system as an indicator of the degree of depression; the longer the period of stillness, the higher the degree of depression. The results are shown below. Figure 6 .

[0294] Depend on Figure 6 It is evident that, compared with the control group, the time spent in a resting state in the Mdivi-1 group was significantly reduced, suggesting that Mdivi-1 can significantly reduce the level of depression after morphine withdrawal.

[0295] 3.5 Mdivi-1 reduces social avoidance levels after spontaneous morphine withdrawal.

[0296] Wild-type mice (i.e., test mice) were randomly divided into the Mdivi-1 group (Mdivi-1) and the control group (Vehicle). Morphine was administered in escalating doses in cages. One hour before each morphine injection, mice were intraperitoneally injected with 50 mg / kg Mdivi-1 (Mdivi-1 group) or an equal volume of corn oil (control group). Nine days after the end of the escalating morphine administration (i.e., nine days of spontaneous morphine withdrawal), a three-box social test was conducted. Each test mouse was first acclimatized to the three-box setup for 3 days, once daily for 10 minutes. The test then began: a strange Stranger 1 mouse (acclimatized to the three-box setup for 3 days, once daily for 10 minutes) was placed on one side of the three boxes, while an empty metal cage was placed on the other side. The test mice were allowed free movement in the three boxes for 10 minutes, and videos were recorded. Video analysis recorded the interaction time between the test mice and the strange Stranger 1 mouse, as well as the time spent exploring the empty cage (including approaching within 2 cm of the cage, sniffing, and direct contact), and the social preference index was calculated. The social index was calculated as: time spent interacting with Stranger 1 mice / time spent exploring the empty cage; a lower index indicated more severe social avoidance. Results are shown in... Figure 7 .

[0297] Depend on Figure 7It is evident that, compared with the control group, the social preference index of mice in the Mdivi-1 group was significantly increased, suggesting that Mdivi-1 can significantly reduce the level of social avoidance after morphine withdrawal.

[0298] Figure 3-7 The results showed that Mdivi-1 could significantly alleviate the physical and psychological symptoms caused by morphine withdrawal, suggesting that Mdivi-1 can prevent or treat physical and psychological dependence caused by long-term morphine use.

[0299] While typical embodiments of the invention have been set forth and described, the invention is not limited to the details described. Various possible modifications and substitutions that do not depart from the spirit of the invention are possible by those skilled in the art through conventional experimentation, and all such modifications and equivalents fall within the spirit and scope of the invention as defined by the following claims.

[0300] sequence list

[0301]

[0302] sequence list <110> Fudan University <120> Uses of mitochondrial division inhibitors and combinations thereof <130> I2021TC5811CS <150> CN202011410107.5 <151> 2020-12-03 <160> 18 <170> PatentIn version 3.5 <210> 1 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Mitochondrial division inhibitory peptide <400> 1 Asp Leu Leu Pro Arg Gly Thr 1 5 <210> 2 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Mitochondrial division inhibitory peptide <400> 2 Asp Leu Leu Pro Arg Gly Ser 1 5 <210> 3 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Membrane-penetrating peptides <400> 3 Arg Arg Arg Gln Arg Arg Lys Lys Arg Gly Tyr 1 5 10 <210> 4 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Membrane-penetrating peptides <400> 4 Arg Arg Gln Arg Arg Thr Ser Lys Leu Met Lys Arg 1 5 10 <210> 5 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Membrane-penetrating peptides <400> 5 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg 1 5 10 <210> 6 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Membrane-penetrating peptides <400> 6 Arg Lys Lys Arg Arg Gln Arg Arg Arg 1 5 <210> 7 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Cell-penetrating peptide <400> 7 Arg Lys Lys Arg Arg Gln Arg Arg 1 5 <210> 8 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Cell-penetrating peptide <400> 8 Tyr Ala Arg Ala Ala Ala Arg Gln Ala Arg Ala 1 5 10 <210> 9 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Cell-penetrating peptide <400> 9 Thr His Arg Leu Pro Arg Arg Arg Arg Arg Arg 1 5 10 <210> 10 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Cell-penetrating peptide <400> 10 Gly Gly Arg Arg Ala Arg Arg Arg Arg Arg Arg 1 5 10 <210> 11 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Cell-penetrating peptide <400> 11 Gly Trp Thr Leu Asn Ser Ala Gly Tyr Leu Leu Gly Lys Ile Asn Leu 1 5 10 15 Lys Ala Leu Ala Ala Leu Ala Lys Lys Ile Leu 20 25 <210> 12 <211> 33 <212> PRT <213> Artificial Sequence <220> <223> Cell-penetrating peptide <400> 12 Lys Ala Leu Ala Trp Glu Ala Lys Leu Ala Lys Ala Leu Ala Lys Ala 1 5 10 15 Leu Ala Lys His Leu Ala Lys Ala Leu Ala Lys Ala Leu Lys Cys Glu 20 25 30 Ala <210> 13 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Cell-penetrating peptide <400> 13 Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys 1 5 10 15 <210> 14 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Membrane-penetrating peptides <400> 14 Arg Arg Arg Arg Arg Arg Arg Arg Arg 1 5 <210> 15 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Polypeptides containing mitochondrial division inhibitory peptides <400> 15 Arg Arg Arg Gln Arg Arg Lys Lys Arg Gly Tyr Gly Gly Asp Leu Leu 1 5 10 15 Pro Arg Gly Ser 20 <210> 16 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Polypeptides containing mitochondrial division inhibitory peptides <400> 16 Arg Arg Arg Gln Arg Arg Lys Lys Arg Gly Tyr Gly Gly Asp Leu Leu 1 5 10 15 Pro Arg Gly Thr 20 <210> 17 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Polypeptides containing mitochondrial division inhibitory peptides <400> 17 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Gly Gly Asp Leu Leu 1 5 10 15 Pro Arg Gly Ser 20 <210> 18 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Polypeptides containing mitochondrial division inhibitory peptides <400> 18 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Gly Gly Asp Leu Leu 1 5 10 15 Pro Arg Gly Thr 20

Claims

1. Use of mitochondrial fission inhibitors in the preparation of medicaments for the prevention or treatment of opioid dependence, wherein the opioid dependence is manifested as physical and psychological dependence; in, The mitochondrial division inhibitor is a compound of formula (II) or a pharmaceutically acceptable salt thereof, wherein the compound of formula (II) has the following structure: (Ⅱ)。 2. The use of claim 1, wherein The drug is used to inhibit the fragmentation of mitochondria in dopaminergic neurons in the ventral tegmental area of ​​the midbrain caused by long-term use of opioid analgesics; or The drug is used to prevent or reduce the subject's dependence on opioid analgesics.

3. A composition for preventing or treating opioid analgesic dependence, said opioid analgesic dependence manifesting as physical and psychological dependence, comprising: (1) The mitochondrial division inhibitor as defined in claim 1 or 2; and (2) Opioid analgesics; Optionally, the composition further comprises a non-opioid analgesic and / or an adjunct analgesic; The non-opioid analgesic is a nonsteroidal anti-inflammatory drug (NSAID); the NSAID is selected from aspirin, indomethacin, acetaminophen, ibuprofen, phenylbutazone, naproxen, naproxen, diclofenac, nimesulide, rofecoxib, celecoxib, ketoibuprofen, flurbiprofen, loxoprofen, zaltoprofen, and combinations thereof; The adjunctive analgesic is selected from corticosteroids, antidepressants, NMDA receptor antagonists, hydroxyzine antihistamines, and combinations thereof.

4. The composition of claim 3, wherein the mitochondrial division inhibitor and the opioid analgesic are present in the same or different compositions.

5. The use of claim 1 or the composition of claim 3 or 4, wherein the opioid analgesic is selected from morphine, dihydromorphine, diacetylmorphine, hydromorphone, hydroxymorphone, oxycodone, hydrocodone, codeine, dihydrocodeine, buprenorphine, methadone, fentanyl, remifentanil, sufentanil, buprenorphine, meperidine, dezocine, butorphanol, bremazocine, carfentanyl, cyclazocine, enadoline, eseroline, ethylmorphine, etonitazine, etorphanine, levofenphene, levofenol, nalbuphine, nicomorphine, pentazocine, phenazocine, picenadol, tramadol, tapentadol, pharmaceutically acceptable salts thereof, and combinations thereof.

6. The use of claim 1 or the composition of claim 3 or 4, wherein the opioid analgesic is selected from morphine, fentanyl, sufentanil, remifentanil, codeine, dihydrocodeine, oxycodone, methadone, pharmaceutically acceptable salts thereof, and combinations thereof.

7. The use of claim 1 or the composition of claim 3 or 4, wherein the opioid analgesic is morphine.

8. The composition of claim 3, wherein the mitochondrial division inhibitor is administered during administration of the opioid analgesic.

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