Cleavage ring C19 diterpene alkaloid compounds and their application in rapid-acting antidepressants

Abstract

This invention relates to the field of pharmaceutical technology, and in particular to a cleaved C19 diterpenoid alkaloid compound and its application in rapid-acting antidepressants. This invention isolates a novel cleaved C19 diterpenoid alkaloid compound, Carmaloidline G, from Aconitum carmichaelii and establishes a stable and controllable extraction, separation, and purification process, achieving high-purity preparation and industrial-scale feasibility of this compound. Studies have shown that this compound exhibits significant rapid-acting antidepressant activity in various animal models of depression, with a fast onset of action and stable efficacy. Its overall efficacy is comparable to the positive control drug esketamine, but the required dosage is significantly reduced, thereby effectively reducing the risk of potential adverse reactions.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to a cleaved C19 diterpenoid alkaloid compound and its application in rapid-acting antidepressants. Background Technology

[0002] In today's increasingly competitive society, the high-intensity, high-pressure lifestyle has a serious impact on people's physical and mental health, leading to a rise in depression and severely affecting people's normal work and life. Currently, most medications for treating depression have significant side effects and, in clinical application, exhibit slow onset of action (such as fluoxetine) and addictive potential (such as esketamine). S Due to limitations such as those of ketamine, there is an urgent need to develop new antidepressants that take effect quickly. Summary of the Invention

[0003] This invention covers the following technical solutions: One aspect of the present invention relates to compounds and their pharmaceutically acceptable salts, solvates or crystal forms, said compounds being cleaved C19 diterpenoid alkaloids having the structure shown in formula (1):

[0004] (1).

[0005] Another aspect of the invention relates to a pharmaceutical composition comprising the compound as described above and its pharmaceutically acceptable salt, solvate or crystal form, and a pharmaceutically acceptable carrier, excipient or excipient.

[0006] Another aspect of the invention relates to the use of the compounds described above and their pharmaceutically acceptable salts, solvates or crystal forms in the preparation of antidepressant drugs.

[0007] Another aspect of the present invention relates to a method for preparing the compound as described above, comprising the following steps: The aconite was extracted with a solvent to obtain the extract, which was then concentrated to obtain the extract paste. The extract is dissolved in an acidic solution, and an acidic aqueous phase is obtained by extraction and separation. The acidic aqueous phase is then alkalized and extracted to obtain an organic phase. The organic phase was then subjected to column chromatography and high-performance liquid chromatography in sequence.

[0008] This invention isolates a novel C19-secreting diterpenoid alkaloid, Carmaloidline G, from Aconitum carmichaelii and establishes a stable and controllable extraction, separation, and purification process, achieving high-purity preparation and industrial-scale feasibility of this compound. Studies have shown that this compound exhibits significant rapid-onset antidepressant activity in various animal models of depression, with a fast onset and stable effect. Its overall efficacy is comparable to the positive control drug esketamine, but the required dosage is significantly lower, effectively reducing the risk of potential adverse reactions. This invention provides a reliable material basis and technical support for the research and development and transformation of novel rapid-onset antidepressants, possessing promising clinical application prospects and industrialization value. Attached Figure Description

[0009] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0010] Figure 1 Compound 1 obtained in Example 1 of this invention 1 1H NMR spectrum (400 MHz, CD3OD); Figure 2 Compound 1 obtained in Example 1 of this invention 13 C NMR spectrum (100 MHz, CD3OD); Figure 3 This is the HMBC spectrum of compound 1 obtained in Example 1 of the present invention. Detailed Implementation

[0011] Reference will now be made to detailed embodiments of the present invention, one or more of which are described below. Each example is provided for explanation and not for limitation of the invention. In fact, it will be apparent to those skilled in the art that various modifications and variations can be made to the invention without departing from its scope or spirit. For example, features described or illustrated as part of one embodiment may be used in another embodiment to produce further embodiments.

[0012] Unless otherwise stated, all terms used to disclose this invention (including technical and scientific terms) should be understood to have the meaning commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein is for the purpose of describing specific embodiments only and is not intended to constitute any limitation on the scope of protection of this invention. Unless the context clearly defines otherwise, the scientific and technical terms used herein, as well as terms and laboratory procedures in related fields such as natural product chemistry, organic chemistry and analytical chemistry, medicinal chemistry, neurobiology, and pharmacology, are all conventional terms and standard methods well-known and widely used in the art. To facilitate understanding of the technical solutions of this invention, some related terms are further defined and explained below.

[0013] As used herein, the terms “and / or,” “or / and,” and “and / or” encompass any one of two or more of the relevant listed items, as well as any and all combinations of the relevant listed items, including any two of the relevant listed items, any more of the relevant listed items, or a combination of all the relevant listed items.

[0014] The terms “containing,” “comprising,” and “including” as used in this invention are synonyms and are inclusive or open-ended, not excluding additional, uncited members, elements, or method steps.

[0015] In this invention, the numerical range represented by endpoints includes all numerical values ​​and fractions contained within that range, as well as the endpoints mentioned.

[0016] Furthermore, in describing representative embodiments of the invention, this specification may present the methods and / or processes of the invention as a specific sequence of steps. However, the method or process should not be limited to the specific order of the steps described herein, to the extent that the method or process does not depend on the specific order of the steps presented herein. As will be understood by those skilled in the art, other sequences of steps are also possible. Therefore, the specific order of steps presented in the specification should not be construed as a limitation of the claims. Additionally, the claims relating to the methods and / or processes of the invention should not be limited to the execution of their steps in the order they are written, and those skilled in the art will readily recognize that the sequence can be changed while still remaining within the spirit and scope of the invention.

[0017] This invention relates to concentration values, which include fluctuations within a certain range. For example, fluctuations are allowed within a corresponding precision range. For instance, 2% can fluctuate within ±0.1%. For larger values ​​or values ​​that do not require overly precise control, even greater fluctuations are permitted.

[0018] As used in this invention, unless otherwise stated, the singular forms of the articles “a,” “an,” and “the” include plural referents.

[0019] In this invention, the terms "multiple" or "various" are used unless otherwise specified, referring to a quantity of 2 or more.

[0020] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.

[0021] In this invention, terms such as "preferred," "better," "more suitable," and "ideal" merely describe implementation methods or embodiments with better effects and should be understood not to limit the scope of protection of this invention. In this invention, terms such as "optionally," "optionally," and "optional" mean that something is optional, that is, selected from either "with" or "without" a parallel solution. If multiple "optional" statements appear in a technical solution, unless otherwise specified and without contradiction or mutual constraint, each "optional" statement is independent.

[0022] As stated herein, “Compound 1,” “C19-type diterpenoid alkaloid compound 1,” and “Carmaloidline G” refer to the same chemical entity, all of which are novel C19-type diterpenoid alkaloid compounds obtained by the present invention through isolation and purification from Aconitum carmichaelii, and have the molecular structure shown in formula (1). In different chapters, examples, or experimental data descriptions, different names are used to refer to the same compound based on writing conventions, ease of expression, or contextual needs, but the objects referred to are completely consistent in terms of chemical structure, physicochemical properties, and pharmacological activity, and all belong to the core active ingredients of the present invention. The use of the above terms in this specification should be understood as equivalent and does not result in any substantial difference due to differences in expression.

[0023] In this invention, the term "cleft-ring C19 diterpenoid alkaloids" refers to a class of natural or semi-natural alkaloid compounds with C19-type diterpenoid alkaloids as the basic skeleton structure, in which some ring systems in the molecule are broken or rearranged to form a cleft-ring structure, and which simultaneously contain nitrogen atoms and multiple oxygen-containing functional groups.

[0024] In this invention, the term "treatment" refers to the process of intervening in a subject's depressive disorder by administering medication. This process includes alleviating, improving, reducing, suppressing, or eliminating depressive symptoms that have already occurred, as well as interventions to prevent, delay, reduce the risk of, or mitigate the potential severity of depressive disorders that have not yet occurred or are at high risk.

[0025] In this invention, the term "pharmaceutically acceptable salt" refers to a salt form formed by the compound of formula (1) with an inorganic acid, organic acid or base that has good biocompatibility and safety in pharmaceutical terms, including but not limited to hydrochloride, sulfate, phosphate, acetate, tartrate, methanesulfonate, citrate and combinations thereof.

[0026] In this invention, the term "solvent" refers to a compound form in which the compound of formula (1) or its pharmaceutically acceptable salt forms a stable binding structure with one or more solvent molecules during crystallization or solidification, including but not limited to hydrates, alcohols and their mixed solvates.

[0027] In this invention, the term "crystal form" refers to the different crystal structures formed by the compound of formula (1) and its pharmaceutically acceptable salts or solvates under solid conditions, which differ in terms of lattice arrangement, physical properties or stability.

[0028] In this invention, the term "pharmaceuticalally acceptable carrier, excipient or excipient" refers to a pharmaceutical excipient component used in a dosage form to improve drug stability, solubility, formability, bioavailability or administration compliance, and which has no significant toxic side effects on the subject.

[0029] In this invention, the term "Aconitum" refers to the processed rootlets of plants of the genus Aconitum in the Ranunculaceae family and their medicinal raw materials, including Aconitum products that have been dried, processed, or pretreated.

[0030] The first aspect of this invention relates to compounds and their pharmaceutically acceptable salts, solvates or crystal forms, said compounds being cleaved C19 diterpenoid alkaloids having the structure shown in formula (1):

[0031] (1).

[0032] The novel split-ring C19 diterpenoid alkaloid compound provided by this invention, with the English name Carmaloidline G, has the molecular structure characteristics shown in formula (1). Structural analysis and mass spectrometry confirmed that the molecular formula of this compound is C19. 24 H 35 NO7, with a relative molecular mass of approximately 449, belongs to the class of small molecule compounds, which are natural products containing nitrogen and multiple functional groups.

[0033] Carmaloidline G typically exists as a yellow powder, exhibiting good chemical stability and exhibiting minimal decomposition or structural degradation under normal temperature and pressure conditions. The compound is readily soluble in polar organic solvents such as methanol and ethanol, but its solubility in water is relatively low, which can be improved by forming pharmaceutically acceptable salts or employing appropriate formulation techniques. This compound demonstrates good physicochemical stability under normal storage conditions, and its purified product maintains a high level of purity, making it suitable as an active pharmaceutical ingredient for further formulation development and efficacy studies. Based on its well-defined molecular formula, stable structural characteristics, and favorable basic physicochemical properties, Carmaloidline G can serve as the core active ingredient in the antidepressant pharmaceutical composition of this invention, providing a reliable material basis for its subsequent pharmaceutical applications and industrialization.

[0034] In some embodiments, the pharmaceutically acceptable salt can be formed by reacting the basic functional group in the Carmaloidline G molecule with inorganic or organic acids, including but not limited to inorganic acid salts such as hydrochloride, sulfate, phosphate, hydrobromide, and nitrate, and organic acid salts such as acetate, tartrate, citrate, oxalate, methanesulfonate, benzenesulfonate, fumarate, succinate, malate, and lactate. These salt forms typically exhibit good water solubility or dissolution properties, which is beneficial for improving the solubility and bioavailability of the active ingredient in the formulation, and for enhancing dosing stability and dosage controllability.

[0035] In some embodiments, the solvate refers to a compound form in which Carmaloidline G or a pharmaceutically acceptable salt thereof forms a stable binding structure with one or more solvent molecules during crystallization, recrystallization, or purification, including but not limited to hydrates, methanol compounds, ethanol compounds, acetone compounds, acetonitrile compounds, or combinations thereof. In the solvate, solvent molecules are embedded in the crystal lattice structure in a stoichiometric or non-stoichiometric manner, thereby affecting the crystal stability, hygroscopicity, flowability, and thermal properties of the compound. In some embodiments of the present invention, preparing the solvate by rationally selecting the solvent system is beneficial to improving the stability of the compound during storage and transportation.

[0036] In some embodiments, the crystal form refers to the different crystal structure arrangements formed by Carmaloidline G and its pharmaceutically acceptable salts or solvates under solid conditions, including polymorphs, metastable crystal forms, amorphous forms, and their interconversion forms. Different crystal forms differ in lattice parameters, molecular packing arrangements, and intermolecular forces, thus exhibiting different characteristics in terms of melting point, dissolution rate, stability, mechanical properties, and bioavailability. In some embodiments of the present invention, different crystal forms can be obtained by controlling process parameters such as solvent type, crystallization temperature, cooling rate, concentration conditions, and drying methods to meet the needs of different dosage form development and administration.

[0037] By systematically screening and optimizing the salt forms, solvates, and crystal forms of Carmaloidline G, comprehensive regulation of its solubility, stability, formulation compatibility, and clinical administration performance can be achieved while maintaining its antidepressant pharmacological activity. This provides a diverse and selectable implementation path for its industrial development as a novel active ingredient in antidepressants.

[0038] A second aspect of the invention relates to a pharmaceutical composition comprising the compound as described above and its pharmaceutically acceptable salt, solvate or crystal form, and a pharmaceutically acceptable carrier, excipient or excipient.

[0039] The pharmaceutical composition can be formulated using methods known to those skilled in the art. For example, it can be administered parenterally in the form of a sterile solution or suspension of water or a pharmaceutically acceptable solution other than water.

[0040] The carrier, excipients, or excipients mentioned in this invention are mainly used to improve the dispersibility, stability, solubility, formability, and administration compliance of the active ingredient, without adversely affecting its pharmacological activity and safety, thereby ensuring the quality controllability and clinical applicability of the drug composition during preparation, storage, and use.

[0041] In some embodiments of the present invention, the pharmaceutically acceptable carrier may be selected from solid carriers, semi-solid carriers, or liquid carriers, including but not limited to lactose, sucrose, glucose, starch, microcrystalline cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyethylene glycol, polyvinylpyrrolidone, dicalcium phosphate, mannitol, sorbitol, vegetable oil, mineral oil, fatty acid glycerides, medium-chain triglycerides, water, or water for injection, etc., to carry and disperse the active ingredient so that it can be uniformly distributed in the formulation system.

[0042] In some embodiments of the present invention, the excipients or excipients may include binders, fillers, disintegrants, lubricants, flow aids, coating agents, solubilizers, emulsifiers, stabilizers, preservatives, antioxidants, buffers, and pH adjusters, etc. Specifically, the binder may be selected from starch paste, gelatin, polyvinylpyrrolidone, or hydroxypropyl cellulose; the filler may be selected from lactose, microcrystalline cellulose, dicalcium phosphate, or mannitol; the disintegrant may be selected from croscarmellose sodium, carboxymethyl starch sodium, or low-substituted hydroxypropyl cellulose; the lubricant and flow aid may be selected from magnesium stearate, talc, or silica; the emulsifier and solubilizer may be selected from Tween derivatives, poloxamer derivatives, or lecithin; and the stabilizer and antioxidant may be selected from sodium bisulfite, vitamin C, tocopherol, or sodium sulfite.

[0043] In some embodiments of the present invention, depending on different routes of administration and dosage form requirements, the pharmaceutically acceptable carriers, excipients, or excipients may further include isotonic modifiers, preservatives, and buffer systems for injectable formulations; viscosity modifiers, sweeteners, and flavoring agents for oral liquids or inhaled formulations; film-forming materials and plasticizers for oral films or sublingual formulations; and lipid materials, surfactants, and stabilizers for liposomes or nanoformulations. Through the rational selection and combination of the types and amounts of the above-mentioned carriers, excipients, or excipients, the pharmaceutical compositions of the present invention can possess good formulation stability, dosing consistency, and clinical applicability while ensuring efficacy and safety, thereby meeting the application needs under different administration routes and treatment scenarios.

[0044] In some embodiments, the dosage form of the pharmaceutical composition is a tablet, capsule, granule, oral liquid, powder, drop, inhaler, oral film, microcapsule, or injection.

[0045] In some embodiments, the pharmaceutical composition is a liposome formulation, a nanoparticle formulation, a microemulsion formulation, or a solid dispersion formulation.

[0046] The chosen route of administration can be rationally selected based on the subject's disease type, disease stage, and clinical medication needs to ensure the stability, bioavailability, and therapeutic effect of Carmaloidline G in vivo.

[0047] A third aspect of the invention relates to the use of the compounds described above and their pharmaceutically acceptable salts, solvates or crystal forms in the preparation of antidepressant medications.

[0048] In some embodiments of the present invention, the antidepressant can be used for long-term treatment, intermittent treatment, or acute intervention for patients with depression, and can also be used for early intervention and preventive medication for individuals at high risk of depression. In some embodiments of the present invention, the antidepressant is used to treat major depressive disorder, mild to moderate depressive disorder, or stress-related depressive disorder. Through rational optimization of the dosing regimen, dosage form structure, and route of administration, the compound can exert stable efficacy in different treatment stages and different patient groups, thereby improving the overall treatment success rate and patients' quality of life.

[0049] The compounds described in this invention exhibited significant and stable antidepressant activity in various animal models of depression, producing clear behavioral improvements within a short time after a single dose, demonstrating excellent rapid onset of action. Experimental results showed that, under the same or similar experimental conditions, the compounds described in this invention, in improving core depressive indicators such as immobility time, spontaneous activity ability, and anhedonia, had an overall efficacy level comparable to that of esketamine, a commonly used rapid-acting positive-positive antidepressant in clinical practice. S It is roughly equivalent to (-Ketamine), but its effective dosage is significantly lower than (-Ketamine). S The compound contains 1-Ketamine, thereby reducing potential adverse reactions and safety risks while ensuring therapeutic efficacy. Based on the above technical effects, the compound of this invention has comprehensive advantages such as rapid onset of action, low dosage, and high safety potential, and can be used as a novel rapid-acting antidepressant candidate for the treatment and intervention of related diseases.

[0050] In some embodiments of the present invention, the compound may also be used in combination with other antidepressants or psychotropic drugs to further enhance the therapeutic effect through synergistic or complementary mechanisms, shorten the onset time, improve the stability of efficacy, and reduce the risk of drug resistance or side effects associated with monotherapy. Other exemplary antidepressants include one or more of the following substances: esketamine (… S Ketamine, fluoxetine, paroxetine, fluvoxamine, sertraline, citalopram, escitalopram, venlafaxine, duloxetine, mirtazapine, bupropion, agomelatine, trazodone, reboxetine, imipramine, amitriptyline, clomipramine, doxepin, maprotiline, moclobemide, liver-soothing and mood-lifting capsules, St. John's wort extract, flupentixol, melitracen.

[0051] The present invention also relates to a method for treating depression, comprising administering to a subject in need the compound as described above and its pharmaceutically acceptable salts, solvates or crystal forms thereof, or administering a pharmaceutical composition comprising the compound and its pharmaceutically acceptable salts, solvates or crystal forms thereof, to relieve, improve or eliminate symptoms associated with depression.

[0052] In some embodiments, the subject of the drug is a mammal. The mammal includes both laboratory animals used for pharmacodynamic, safety, and mechanistic studies, and non-laboratory animals used in clinical or veterinary settings for disease prevention or treatment. By applying Carmaloidline G to different mammalian subjects, its pharmacological effects, administration methods, and therapeutic efficacy can be systematically evaluated, thereby validating its applicability and universality in depression intervention. Exemplary mammals include, but are not limited to, humans, as well as non-human mammals such as mice, rats, guinea pigs, hamsters, rabbits, dogs, cats, pigs, sheep, cattle, pandas, elephants, horses, and non-human primates. More preferably, the subject is a human.

[0053] In some embodiments of the present invention, the compounds described above and their pharmaceutically acceptable salts, solvates, or crystal forms can be rationally administered according to factors such as the subject's age, weight, disease severity, route of administration, and individual tolerance. The specific dosage can be flexibly adjusted while ensuring efficacy and safety. Generally, the daily dosage of the compounds can be 0.001–10 mg / kg, preferably 0.01–1 mg / kg, and more preferably 0.03–0.5 mg / kg. This dosage range can cover different clinical needs for low-dose onset of action and medium-to-high-dose maintenance therapy.

[0054] In some embodiments of the present invention, the Carmaloidline G and its pharmaceutically acceptable salts, solvates, or crystal forms can be administered to subjects via various routes of administration, including but not limited to oral administration, injection administration, inhalation administration, nasal administration, sublingual administration, or oral mucosal administration. Oral administration is suitable for long-term maintenance therapy and outpatient medication management; injection administration is suitable for acute exacerbations or clinical situations requiring rapid onset of action; inhalation, nasal, or sublingual administration can be used to improve the onset of action and bioavailability, and reduce the impact of the first-pass effect on efficacy.

[0055] In some embodiments of the present invention, the compound may be designed as an immediate-release formulation, a sustained-release formulation, or a controlled-release formulation, depending on the route of administration and dosage form. Immediate-release formulations are suitable for acute treatment phases requiring rapid symptom relief, while sustained-release or controlled-release formulations are suitable for stable or maintenance treatment phases, achieving a steady release of drug concentrations in the body, thereby reducing the frequency of administration and improving patient compliance.

[0056] In some embodiments of the present invention, the treatment course can be individually adjusted according to the patient's specific condition and clinical response. Generally, the acute phase treatment course can be 1 to 4 weeks, used to rapidly improve the core symptoms of depression; the consolidation phase treatment course can be 4 to 12 weeks, used to stabilize the therapeutic effect and prevent relapse; the maintenance phase treatment course can be 3 to 12 months or longer, used to reduce the risk of relapse and improve long-term prognosis. In some cases requiring rapid intervention, a single-dose or short-course dosing regimen can also be used to obtain a rapidly effective treatment.

[0057] In some embodiments of the present invention, the compound may be used alone or in combination with other antidepressants to further improve therapeutic efficacy or reduce the risk of adverse reactions from high doses of a single drug. In cases of combination therapy, the dosage, dosing sequence, and dosing interval can be rationally designed based on the different mechanisms of action and pharmacokinetic characteristics of the drugs to achieve synergistic effects and safe medication use.

[0058] This invention also relates to a method for preparing the compound as described above, comprising the following steps: The aconite was extracted with a solvent to obtain the extract, which was then concentrated to obtain the extract paste. The extract is dissolved in an acidic solution, and an acidic aqueous phase is obtained by extraction and separation. The acidic aqueous phase is then alkalized and extracted to obtain an organic phase. The organic phase was then subjected to column chromatography and high-performance liquid chromatography in sequence.

[0059] In some embodiments, the solvent is one or more of methanol, ethanol, acetone, chloroform, or petroleum ether.

[0060] In some embodiments, the solvent is an aqueous ethanol solution with a volume fraction of 88% to 98%, the amount of solvent used is 8 to 10 times the mass of the plant raw material, and the extraction is carried out by reflux 2 to 4 times, each time for 1 to 3 hours.

[0061] In some embodiments, the acidic solution is dilute hydrochloric acid with a mass fraction of 0.1% to 1.0%.

[0062] In some embodiments, the pH of the acidic aqueous phase after alkalization is 8 to 11.

[0063] In some embodiments, the high-performance liquid chromatography mobile phase is an acetonitrile-water system with a volume ratio of (26-30):(70-74) and contains 0.01%-0.5% trifluoroacetic acid.

[0064] In some embodiments, the high-performance liquid chromatography mobile phase is an acetonitrile-water system with a volume ratio of 28:72, and contains 0.1% trifluoroacetic acid.

[0065] In some embodiments, the high-performance liquid chromatography uses a C18 column.

[0066] In some embodiments, the column chromatography separation sequentially includes silica gel column chromatography, ODS column chromatography, and gel column chromatography (preferably Sephadex LH-20 gel column chromatography). More specifically, the column chromatography separation includes: S1. The extract B was subjected to gradient elution using a mobile phase via silica gel column chromatography to obtain fraction C; when identified by silica gel thin-layer chromatography, the Rf value corresponding to fraction C was 0.72-0.75. S2. Using a mobile phase of methanol-water, fraction C was eluted by ODS column chromatography to obtain fraction C3; when identified by silica gel thin-layer chromatography, the Rf value corresponding to fraction C3 was 0.50-0.60. S3. Using a mobile phase of methanol-water, fraction C3 was subjected to gel column chromatography to obtain fraction M1; the Rf value of fraction M1 was 0.30-0.39. S4. The fraction M1 was subjected to reversed-phase high-performance liquid chromatography with acetonitrile-water as the mobile phase to obtain a novel split-ring C19 diterpenoid alkaloid compound Carmaloidline G with the structural formula shown in formula (1). The preferred retention time of the compound Carmaloidline G is 21-23 min.

[0067] In some embodiments, based on a total volume of 100, in step S1, the silica gel column chromatography employs gradient elution, and the mobile phase consists of solvent A and solvent B in a volume ratio of (0:100)-(100:0); preferably (93:7)-(91:9), more preferably a mixed solvent of 92:8. Solvent A is any one of dichloromethane, chloroform, and petroleum ether; Solvent B is any one of chloroform, acetone, ethyl acetate, and methanol.

[0068] In some more specific embodiments, the silica gel column chromatography employs a dichloromethane-methanol system for gradient elution.

[0069] In some embodiments, the ODS column chromatography uses a methanol-water system as the mobile phase, and contains 0.01% to 0.1% (preferably 0.05%) formic acid. More preferably, the volume ratio of methanol to water in the methanol-water system is (35:65) to (45:55); more preferably 40:60.

[0070] In some embodiments, the Sephadex LH-20 gel column chromatography uses a methanol-water system as the mobile phase. More preferably, the volume ratio of methanol to water in the methanol-water system is (1:100) to (10:90); more preferably, it is 5:95.

[0071] The embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that these embodiments are only used to illustrate the technical content of the present invention and are not intended to limit the scope of protection of the present invention. Unless otherwise specified, the specific experimental conditions in the following embodiments are given priority reference to the guidelines provided in this specification, or may be carried out according to generally accepted experimental manuals or conventional experimental conditions, or other experimental methods known in the art, or according to the conditions recommended by the relevant reagent or instrument manufacturers. In specific embodiments, unless otherwise specified, minor deviations within the weighing accuracy range are allowed for the measurement parameters involving raw material components; reasonable deviations due to instrument detection accuracy or operational accuracy are also allowed for parameters such as temperature and time.

[0072] Example 1 A method for preparing a novel type of split-ring C19 diterpenoid alkaloid compound, Carmaloidline G, includes the following steps: S1. Take dried Aconitum carmichaelii (about 200.5 kg), use 95% ethanol aqueous solution (10 times the weight of Aconitum carmichaelii) as solvent for reflux extraction 3 times, each extraction for 2 hours, combine the extracts and concentrate to obtain extract (about 6 kg). S2. Add the extract to 10 times its mass of 0.5% dilute hydrochloric acid (60L) to fully dissolve and suspend it. Filter to remove impurities. Extract the filtered acidic aqueous solution three times with 1.5 times its volume of dichloromethane to obtain acidified dichloromethane extract A (about 2.5kg, discarded) and acidic aqueous solution A (about 1.2kg).

[0073] The pH of the acidic aqueous solution A was adjusted to 9 with sodium hydroxide solution, and then extracted with dichloromethane to obtain the dichloromethane extract, namely extract B (about 500g).

[0074] S3. The extract B was subjected to column chromatography and high performance liquid chromatography in sequence to obtain a novel cleaved C19 diterpenoid alkaloid compound.

[0075] Step S3 specifically includes the following steps: S31. Extract B is subjected to gradient elution using dichloromethane-methanol silica gel column chromatography to obtain fraction C; when identified by silica gel thin-layer chromatography, the Rf value corresponding to fraction C is 0.72-0.75. The mobile phase for silica gel column chromatography is dichloromethane-methanol with a volume ratio of (93:7)-(91:9). Preferably, the volume ratio of dichloromethane-methanol in the mobile phase is 92:8.

[0076] It should be noted that during gradient elution, dichloromethane-methanol at volume ratios of (93:7) to (91:9) can elute fraction C. Using silica gel thin-layer chromatography, fractions with an Rf value of 0.72-0.75 are identified as fraction C. The dichloromethane-methanol volume ratio of 92:8 exhibits the highest elution efficiency. The principle behind the "preferred volume ratio" described in the following gradient elution processes is the same.

[0077] Specifically, extract B was subjected to silica gel column chromatography, eluted with a dichloromethane-methanol gradient at volume ratios of 100:0, 95:5, 92:8, 90:10, 85:15, 75:25, 65:35, 50:50, and 0:100, yielding fractions 1-10, 11-20, 21-30, 31-40, 41-50, 51-60, 61-70, 71-80, and 81-90, for a total of 90 fractions. These fractions were then identified using silica gel thin-layer chromatography. Based on the brick-red spots observed by potassium bismuth iodide reagent, the Rf values ​​were 0.82-0.86 (fractions 1-10) and 0.76-0.80 (fractions 1-10). Fractions were divided into four groups: 0.72-0.75 (fraction 21-30), 0.65-0.70 (fraction 31-40), 0.56-0.62 (fraction 41-50), 0.50-0.54 (fraction 51-60), 0.42-0.48 (fraction 61-70), 0.30-0.38 (fraction 71-80), and 0.25-0.28 (fraction 81-90). Similar fractions were combined to obtain fractions A, B, C, D, E, F, G, H, and I. Based on the brick-red spots observed by thin-layer chromatography and the characteristic ultraviolet absorption (λ) of the secoirous diterpenoid alkaloids observed by HPLC analysis, the presence of potassium bismuth iodide was determined. max =252nm) Fraction C was selected for the next separation step. Among them, fractions 21-30 were combined to obtain fraction C.

[0078] S32. Eluent fraction C using a methanol-water mobile phase via ODS column chromatography to obtain fraction C3; when identified by silica gel thin-layer chromatography, the Rf value corresponding to fraction C3 is 0.50-0.60. The volume ratio of methanol-water in the ODS column chromatography mobile phase is (35:65)-(45:55), preferably 40:60. The mobile phase also contains 0.01-0.1% formic acid by volume, preferably 0.05% formic acid by volume.

[0079] The specific operation is as follows: fraction C is subjected to ODS column chromatography and eluted with a methanol-water gradient with volume ratios of 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, and 100:0. The mobile phase methanol-water also contains 0.05% formic acid by volume.

[0080] Fractions 1-5, 6-15, 16-25, 26-35, 36-45, 46-50, 51-55, 56-60, and 61-68 were obtained sequentially, totaling 68 fractions. These were identified using silica gel thin-layer chromatography. Based on the brick-red spots observed with potassium bismuth iodide reagent, the Rf values ​​were 0.35-0.44 (fractions 1-5), 0.45-0.49 (fractions 6-15), 0.50-0.60 (fractions 16-25), 0.61-0.69 (fractions 26-35), 0.70-0.74 (fractions 36-55), 0.75-0.79 (fractions 56-60), and 0.80-0.89 (fractions 61-68). Similar fractions were combined into seven fractions, C1-C7. Fraction 16-25 was combined to obtain fraction C3.

[0081] Based on the brick-red spots observed by potassium bismuth iodide reagent in thin-layer chromatography and the characteristic ultraviolet absorption (λ) of cleaved diterpenoid alkaloids observed by HPLC analysis, [further details are needed]. max =252nm) Select fraction C3 for the next separation step.

[0082] S33. Fraction C3 is subjected to gel column chromatography using a mobile phase of methanol-water to obtain fraction M1; the Rf value of fraction M1 is 0.30-0.39; the volume ratio of methanol-water in the gel column chromatography mobile phase is (0:100)-(10:90). Preferably, the volume ratio of methanol-water is 5:95.

[0083] The specific procedure included: fraction C3 was subjected to dextran gel LH-20 column chromatography, using a methanol-water gradient elution with volume ratios of 5:95, 30:70, 70:30, and 100:0, collecting fractions 1-6, 7-14, 15-20, and 21-29 sequentially, for a total of 29 fractions. These fractions were then identified using silica gel thin-layer chromatography. Based on the brick-red spots observed by potassium bismuth iodide reagent, the Rf values ​​were 0.30-0.39 (fractions 1-6), 0.40-0.54 (fractions 7-14), 0.55-0.61 (fractions 15-20), and 0.62-0.85 (fractions 21-29), respectively. Similar fractions were combined to obtain four fractions M1 to M4. Fractions 1-6 were combined to obtain fraction M1.

[0084] Based on the brick-red spots observed by potassium bismuth iodide reagent and the characteristic ultraviolet absorption (λ) of cleaved diterpenoid alkaloids observed by HPLC analysis. max =252nm) Select fraction M1 for the next separation step.

[0085] S34. The fraction M1 was subjected to reversed-phase high-performance liquid chromatography. The mobile phase had a volume ratio of acetonitrile-water of (26:74)-(30:70) and also contained 0.01-0.5% trifluoroacetic acid by volume. A novel split-ring C19 diterpenoid alkaloid compound Carmaloidline G with the structural formula shown in formula (1) was obtained. The retention time of compound Carmaloidline G was 21-23 min.

[0086] The specific procedure was as follows: HPLC was used with acetonitrile-water (28:72 v / v) as the mobile phase, which also contained 0.1% trifluoroacetic acid (v / v). A C18 column was used to prepare a C19-type diterpenoid alkaloid compound 1 (i.e., compound Carmaloidline G) in fraction M1. t R =22min, purity is 98%).

[0087] The physical properties and detection data of compound 1 obtained in Example 1 are as follows: Yellow powder, readily soluble in methanol. High-resolution mass spectrometry (HR-ESI-MS) m / z 450.2484 ([M+H] + The calculated value is 450.2486, and its molecular formula is determined to be C. 24 H 35 NO7 has an unsaturation degree of 8.

[0088] 1 1H NMR (CD3OD, 400MHz) spectrum Figure 1 (This indicates a signal for one olefin proton) d H 6.93 (1H, t, J = 2.3Hz); 19 proton signals on carbon atoms bonded to oxygen / nitrogen [ d H 4.14 (1H, d, J = 3.9 Hz), 4.04 (1H, dd, J =5.8, 2.5 Hz), 3.81 (1H, s), 3.53 (2H, abq, J = 12.8, 8.5 Hz), 3.39 (3H, s), 3.33 (6H, overlapped), 3.26 (1H, overlapped), 3.22 (1H, overlapped), 3.16 (2H, abq, J = 12.6 Hz); hydrogen atoms on the three adjacent carbons of the double bond [ dH 3.24 (1H, overlapped), 3.07 (1H,d, J = 2.5 Hz), 2.98 (1H, m)] and d H The signal values ​​for one methyl group, three methylene groups, and one methine proton were between 2.50 and 1.25. The proton NMR spectrum suggests that compound 1 may be an amino alcohol-type C19 diterpenoid alkaloid with a double bond.

[0089] 13 C NMR (CD3OD, 100MHz) spectrum Figure 2 The spectrum showed a total of 24 carbon signals, and based on the HSQC spectrum, it was determined that the compound contained two carbonyl carbon signals. d C 193.7 (C-8), 176.9 (C-15)]; a set of olefin carbon signals [ d C 145.5 (C-9), 144.0 (C-14)]; 10 carbon signals linked to oxygen / nitrogen [ d C 84.5 (C-6), 78.9 (C-18), 72.4 (C-16), 69.3 (C-1), 64.8 (C-17), 59.4 (18-OCH3), 59.0 (6-OCH3), 57.9 (C-19), 50.6 (C-20), 49.9 (16-OCH3)]; 1 methyl carbon signal d C 10.4 (C-21); 3 methylene carbon signals [ d C 34.3 (C-12), 29.9 (C-2), 29.2 (C-3)]; 4 methylene carbon signals [ d C 54.7 (C-7), 52.6 (C-10), 49.6 (C-13), 47.4 (C-5)] and two quaternary carbon signals [ d C [50.6 (C-11), 47.4 (C-5)]. Based on the above information, it is speculated that compound 1 may be a cleaved C19 type diterpenoid alkaloid.

[0090] 1 H NMR, 13 The signal assignments for C NMR are shown in Table 1.

[0091] Table 1 1 H NMR, 13Signal attribution of C NMR d (ppm)

[0092] In HMBC spectrum ( Figure 3 In ), H-16 ( d H 4.14) and C-15 ( d C 176.9), C-13 ( d C 49.6) and C-12 ( d C 34.3) has remote correlation, H-14 ( d H 6.93) and C-8 ( d C 193.7) and C-10 ( d C 52.6) has remote correlation and H-6 ( d H 4.04) and C-8 ( d C 193.7) exhibits long-range correlation, and combined with chemical shift and molecular formula, indicates that this compound is a C-8 / C-15 oxidatively cleaved C-8 / C-15 compound. 19 It is a type of diterpenoid alkaloid with the C-8 position oxidized to a ketone carbonyl group and the C-15 position oxidized to a carboxylic acid group. 16-OCH3 ( d H 3.33) and C-16 (δ C 72.4) has a long-range correlation, indicating that the C-16 position of compound 1 is methoxy-substituted.

[0093] In the ROESY spectrum, H-1 ( d H 4.40) and H-12 α ( d H 2.43) related signals and H-12 β ( d H 2.20) and H-5 ( d H The correlation signal (2.40) indicates that the proton at the C-1 position is... α Orientation. H-6 ( d H 4.04) and H-10 ( d H The correlation signal (2.98) indicates that the proton at position C-6 is... β Orientation.

[0094] The CD spectrum of compound 1 is shown at 212 (Δ e -0.62) nm and 236 (Δ e There is a negative Cotton effect at -0.52 nm and at 293 (Δ e There is a positive Cotton effect at 0.65 nm. After derivatization, the Cotton effect changes, and it changes to 296 (Δ) nm. e There is a negative Cotton effect at -1.19 nm and at 347 (Δ e A positive Cotton effect exists at 0.31 nm. This is achieved by comparing with... S Comparison of the CD spectra after derivatization with (+)-lactic acid confirmed that the hydroxyl group at C-15 was... S Configuration.

[0095] In summary, the structure of the new compound 1 (compound Carmaloidline G) was determined as follows:

[0096] Comparative Example 1 The preparation method is the same as in Example 1, except that in step S34, the aqueous solution of the mobile phase used to prepare the mobile phase for separating fraction M1 by HPLC does not contain trifluoroacetic acid.

[0097] The compound prepared in Comparative Example 1 had severe peak tailing during the preparation process, and liquid chromatography analysis after preparation revealed a large number of impurities. The purity of the prepared compound 1 was less than 50%.

[0098] Comparative Example 2 The preparation method is the same as that in Example 1, except that in step S32, when fraction C is subjected to ODS column chromatography, methanol-water with an initial volume ratio of 40:60 is used for elution, and the mobile phase does not contain formic acid.

[0099] In Comparative Example 2, this compound could not be detected during subsequent fraction preparation. The results showed that compound 1 could not be prepared.

[0100] Experimental Example 1: Carmaloidline G-based mouse behavioral despair model experiment 1. Experimental materials and instruments Escloramine was purchased from Jiangsu Hengrui Medicine Co., Ltd.; Higenamine, Bulleyaconitine A, Lappaconitine, and 3-Acetylaconitine were purchased from Shanghai Standard Technology Service Co., Ltd. Songorine, total alkaloids of Aconitum carmichaeli Debx, and Carmaloidline G were prepared in the laboratory.

[0101] ZIL-2 type mouse spontaneous activity box (Shanghai Xinman Science and Education Equipment Co., Ltd.); YLS-18A type mouse tail suspension apparatus (Anhui Zhenghua Biological Instrument Equipment Co., Ltd.); electronic balance (Sartorius Scientific Instruments (Beijing) Co., Ltd.).

[0102] 2. Experimental animals C57BL / 6J male mice, weighing 18 - 22 g, SPF grade, provided by Vital River Laboratory Animal Technology Co., Ltd., animal license number: SYXK (Beijing) 2023 - 0001. The animals were housed in an environment with a temperature of 23 ± 2°C and a humidity of 50 ± 10%, and the lighting time was 12 h per day (7:00 - 19:00 for lighting). After 3 days of animal feeding, various experiments were carried out. The animals were fasted for 6 h before the start of the behavioral experiments and had free access to water.

[0103] 3. Experimental methods 100 C57BL / 6J male mice were arranged in a snake-like pattern and divided into 10 groups according to the descending order of the spontaneous activity times during animal screening, namely the blank group (normal saline of the same volume), positive drug ( S )-Ketamine group (10 mg / kg), Higenamine group (10 mg / kg), Bulleyaconitine A group (10 mg / kg), Lappaconitine group (10 mg / kg), 3-Acetylaconitine group (10 mg / kg), Songorine group (10 mg / kg), total alkaloids of Aconitum carmichaeli Debx group (10 mg / kg), Carmaloidline G low-dose group (0.03 mg / kg), Carmaloidline G high-dose group (0.3 mg / kg), with 10 mice in each group, and they were numbered separately. The drugs were all dissolved in normal saline to prepare solutions, and the administration volume was 0.1 ml / 10 g body weight. The tail suspension experiment and the spontaneous activity experiment were carried out 1 h after a single intraperitoneal injection.

[0104] 4. Experimental results As shown in Table 2, 1 hour after a single intraperitoneal injection, compared with the blank group, the various dose groups of Carmaloidline G (0.03, 0.3 mg / kg) significantly shortened the immobility time of mice in the tail suspension experiment, showing significant rapid antidepressant activity. It was significantly better than norcodonine (10 mg / kg), aconitine (10 mg / kg), eucommiaine (10 mg / kg), 3-acetyl aconitine (10 mg / kg), succinate (10 mg / kg) and total alkaloids of Aconitum carmichaelii (10 mg / kg), and comparable to the positive control drug esketamine (10 mg / kg, a first-line drug for rapid clinical treatment of depression).

[0105] Table 2. Effect of Carmaloidline G on tail suspension immobility time in mice with a behavioral despair model.

[0106] (Compared with the blank group, P <0.05, P <0.0001) As shown in Table 3, compared with the blank group, none of the drug administration groups had a significant effect on the spontaneous activity of mice in the open field experiment, indicating that the effect of each drug administration group on the immobility time of mice in the tail suspension experiment was not related to nerve excitability.

[0107] Table 3. Effects of Carmaloidline G on open field horizontal movement distance in mice of the behavioral despair model

[0108] Experimental Example 2: Carmaloidline G Corticosterone-Induced Mouse Depression Model Experiment (Drug-Induced Model) 1. Experimental materials and instruments Same as item 1 under Experiment 1.

[0109] 2. Laboratory animals Same as Example 1, item 2.

[0110] 3. Experimental Methods One hundred and twenty male C57BL / 6J mice were randomly divided into 12 groups in descending order of spontaneous activity, arranged in a serpentine pattern: a control group (same volume of physiological saline), a model group (corticosterone 40 mg / kg), a positive control group (esketamine 10 mg / kg), aconitine group (10 mg / kg), aconitine group (10 mg / kg), aconitine-rich group (10 mg / kg), 3-acetyl aconitine group (10 mg / kg), succinate group (10 mg / kg), total alkaloids of Aconitum carmichaelii group (10 mg / kg), low-dose Carmaloidline G group (0.03 mg / kg), medium-dose Carmaloidline G group (0.1 mg / kg), and high-dose Carmaloidline G group (0.3 mg / kg). Each group consisted of ten mice, each numbered. All drugs were dissolved in physiological saline at a dose of 0.1 ml / 10 g body weight. Except for the control group, all other groups received intraperitoneal injections of corticosterone 40 mg / kg for 21 consecutive days. On day 22, spontaneous activity test and tail suspension test were performed 1 hour after a single intraperitoneal injection. On day 23, sucrose preference test was performed 1 hour after a single intraperitoneal injection.

[0111] 4. Experimental Results As shown in Table 4, 1 hour after a single intraperitoneal injection, compared with the model group, the various dose groups of Carmaloidline G (0.03, 0.1, and 0.3 mg / kg) significantly shortened the immobility time of mice in the tail suspension experiment and reversed the behavioral despair induced by corticosterone in depressed mice. They were significantly superior to norcodonine (10 mg / kg), aconitine (10 mg / kg), eucommiaine (10 mg / kg), 3-acetyl aconitine (10 mg / kg), succinate (10 mg / kg), and total alkaloids of Aconitum carmichaelii (10 mg / kg), and comparable to the positive control drug esketamine (10 mg / kg).

[0112] Table 4. Effect of Carmaloidline G on tail immobility time in corticosterone-induced mouse model

[0113] (Compared with the blank group, #### P <0.0001; compared with the model group, P <0.05, P <0.01, P <0.001, P <0.0001) As shown in Table 5, 1 hour after a single intraperitoneal injection, compared with the model group, the medium and high dose groups of Carmaloidline G (0.1 and 0.3 mg / kg) significantly increased the number of horizontal traversal grids in mice in the open field experiment and improved the motor ability of corticosterone-induced depressed mice. They were significantly better than norcodonine (10 mg / kg), aconitine (10 mg / kg), aconitine (10 mg / kg), 3-acetyl aconitine (10 mg / kg), succinate (10 mg / kg), and total alkaloids of Aconitum carmichaelii (10 mg / kg), and comparable to the positive control drug esketamine (10 mg / kg).

[0114] Table 5. Effect of Carmaloidline G on the number of horizontal traversing grids in a corticosterone-induced mouse model.

[0115] (Compared with the blank group, #### P <0.0001; compared with the model group, P <0.05, P <0.01) As shown in Table 6, 1 hour after a single intraperitoneal injection, compared with the model group, the medium and high dose groups of Carmaloidline G (0.1 and 0.3 mg / kg) significantly increased the number of paw lifts in mice in the open field experiment and enhanced the exploration desire of corticosterone-induced depressed mice. This was significantly better than norcodonine (10 mg / kg), aconitine (10 mg / kg), aconitine (10 mg / kg), 3-acetyl aconitine (10 mg / kg), succinate (10 mg / kg), and total alkaloids of Aconitum carmichaelii (10 mg / kg), and comparable to the positive control drug esketamine (10 mg / kg).

[0116] Table 6. Effect of Carmaloidline G on the number of foot lifts induced by corticosterone in a mouse model.

[0117] (Compared with the blank group, #### P <0.0001; compared with the model group, P <0.05, P <0.001) As shown in Table 7, 1 hour after a single intraperitoneal injection, compared with the model group, the medium and high dose groups of Carmaloidline G (0.1 and 0.3 mg / kg) significantly increased the sucrose preference index of mice in the sucrose preference experiment and improved the anhedonia induced by corticosterone in depressed mice. They were significantly better than norcodonol (10 mg / kg), aconitine (10 mg / kg), eucommia (10 mg / kg), 3-acetyl aconitine (10 mg / kg), succinyl alkaloids (10 mg / kg) and total alkaloids of Aconitum carmichaelii (10 mg / kg), and comparable to the positive control drug esketamine (10 mg / kg).

[0118] Table 7. Effects of Carmaloidline G on sucrose preference in a corticosterone-induced mouse model.

[0119] (Compared with the blank group, ### P <0.001; compared with the model group, P <0.05, P <0.001) Experiment Example 3: Carmaloidline G Chronic Restraint Stress (CRS) Induced Mouse Depression Model Experiment (Chronic Depression Model) 1. Experimental materials and instruments Same as item 1 under Experiment 1.

[0120] 2. Laboratory animals Same as Example 1, item 2.

[0121] 3. Experimental Methods One hundred and ten male C57BL / 6J mice were randomly divided into 11 groups in a serpentine pattern, ordered by the number of spontaneous activities during screening: control group, model group, esketamine positive control group (10 mg / kg), norcodonol group (10 mg / kg), aconitine group (10 mg / kg), high aconitine group (10 mg / kg), 3-acetylaconitine group (10 mg / kg), succinate group (10 mg / kg), total aconitine group (10 mg / kg), low-dose carmaloidline G group (0.1 mg / kg), and high-dose carmaloidline G group (0.3 mg / kg). All drugs were prepared by dissolving them in physiological saline at a dose of 0.1 ml / 10 g body weight. Except for the control group, all other groups underwent chronic restraint stress for 4 hours daily for 21 consecutive days. On day 22, spontaneous activity and tail suspension tests were performed 1 hour after a single intraperitoneal injection. On day 23, sucrose preference tests were performed 1 hour after a single intraperitoneal injection.

[0122] 4. Experimental Results As shown in Table 8, 1 hour after a single intraperitoneal injection, compared with the model group, the various dose groups of Carmaloidline G (0.1, 0.3 mg / kg) significantly shortened the immobility time of mice in the tail suspension experiment and reversed the behavioral despair induced by CRS in depressed mice. They were significantly superior to norcodonine (10 mg / kg), aconitine (10 mg / kg), eucommiaine (10 mg / kg), 3-acetyl aconitine (10 mg / kg), succinyl alkaloids (10 mg / kg) and total alkaloids of Aconitum carmichaelii (10 mg / kg), and comparable to the positive control drug esketamine (10 mg / kg).

[0123] Table 8. Effect of Carmaloidline G on tail suspension immobility time in CRS-induced mouse model

[0124] (Compared with the blank group, ## P <0.01; compared with the model group, P <0.05, P <0.01, P <0.001) As shown in Table 9, 1 hour after a single intraperitoneal injection, compared with the model group, each dose group of Carmaloidline G (0.1, 0.3 mg / kg) significantly increased the number of horizontal traversal grids in mice in the open field experiment and improved the motor ability of CRS-induced depressed mice. It was significantly better than norcodonine (10 mg / kg), aconitine (10 mg / kg), eucommiaine (10 mg / kg), 3-acetyl aconitine (10 mg / kg), succinyl alkaloids (10 mg / kg), total alkaloids of Aconitum carmichaelii (10 mg / kg), and the positive control drug esketamine (10 mg / kg).

[0125] Table 9. Effect of Carmaloidline G on the number of horizontal traversing grids in a CRS-induced mouse model.

[0126] (Compared with the blank group, #### P <0.0001; compared with the model group, P <0.05, P <0.001) As shown in Table 10, 1 hour after a single intraperitoneal injection, compared with the model group, the various dose groups of Carmaloidline G (0.1, 0.3 mg / kg) significantly increased the number of paw lifts in mice in the open field experiment and enhanced the exploration desire of CRS-induced depressed mice. They were significantly better than norcodonine (10 mg / kg), aconitine (10 mg / kg), eucommiaine (10 mg / kg), 3-acetyl aconitine (10 mg / kg), succinyl alkaloids (10 mg / kg) and total aconite alkaloids (10 mg / kg), and comparable to the positive control drug esketamine (10 mg / kg).

[0127] Table 10 Effect of Carmaloidline G on the number of foot lifts in a CRS-induced mouse model

[0128] (Compared with the blank group, # P <0.05; compared with the model group, P <0.05, P <0.01, P <0.001) As shown in Table 11, 1 hour after a single intraperitoneal injection, compared with the model group, Carmaloidline G (0.1 mg / kg) significantly increased the sucrose preference index in mice in the sucrose preference experiment and improved the anhedonia induced by CRS in depressed mice. It was significantly better than norcodonine (10 mg / kg), aconitine (10 mg / kg), eucommia ulmoides (10 mg / kg), 3-acetyl aconitine (10 mg / kg), succinyl alkaloids (10 mg / kg) and total alkaloids of Aconitum carmichaelii (10 mg / kg), and comparable to the positive control drug esketamine (10 mg / kg).

[0129] Table 11 Effects of Carmaloidline G on sucrose preference in CRS-induced mouse models

[0130] (Compared with the blank group, #### P <0.0001; compared with the model group, P <0.05, P <0.01) In summary, the novel cleaved C19 diterpenoid alkaloid compound Carmaloidline G described in this invention exhibits significant rapid-onset antidepressant activity, comparable to esketamine, a first-line drug for the rapid clinical treatment of depression. S It is comparable to (-Ketamine), but the dosage is much smaller. S )-Ketamine is a potential candidate for a fast-acting antidepressant.

[0131] Application Example 1 This invention discloses a capsule formulation using Carmaloidline G as the active pharmaceutical ingredient, the components of which are as follows: Carmaloidline G 3.5mg 6.0g of starch Sodium metabisulfite 0.2g Magnesium stearate 0.2g Anhydrous ethanol appropriate amount Make 100 pills.

[0132] The specific preparation process is as follows: Carmaloidline G, starch, and sodium metabisulfite are mixed evenly, and anhydrous ethanol is added to make a soft material. The mixture is then passed through a 24-mesh sieve to form granules, dried, and magnesium stearate is added. The mixture is then mixed and filled into capsules.

[0133] Application Example 2 This invention discloses a granule formulation using the compound Carmaloidline G as a raw material, the components of which are as follows: Carmaloidline G 4.0mg 6.0g of starch Sodium bisulfite 0.2g Magnesium stearate 0.2g Anhydrous ethanol appropriate amount Make 100 bags.

[0134] The specific preparation process is as follows: Carmaloidline G was mixed with starch and sodium bisulfite, and then anhydrous ethanol was added to make a soft material. The material was passed through a 24-mesh sieve to make granules, dried, and then magnesium stearate was added. The mixture was then mixed and packaged.

[0135] Application Example 3 This invention discloses an oral liquid using the compound Carmaloidline G as a raw material, the components of which are as follows: Carmaloidline G 6.0mg 3.0g sucrose Sodium bisulfite 0.2g Methylparaben 0.2g Sodium bicarbonate 0.5mg 1000.0 mL of water for injection Make 100 pieces.

[0136] The specific preparation process is as follows: After the above components are mixed, they can be dispensed using conventional oral liquid preparation methods.

[0137] Application Example 4 This invention discloses an injectable formulation using the compound Carmaloidline G as a raw material, the components of which are as follows: Carmaloidline G 4.0mg Vitamin C 0.2g Sodium chloride 6.0g Sodium bicarbonate 0.5mg 1000.0 mL of water for injection Make 100 pieces.

[0138] The specific preparation process is as follows: After the above components are mixed, 100 vials can be obtained by using conventional injection preparation methods.

[0139] Application Example 5 This invention discloses an application example of a compound Carmaloidline G and esketamine (( S Tablets containing ketamine as the active pharmaceutical ingredient have the following components: Carmaloidline G 3.5mg Esketamine 10.0mg Hydroxypropyl methylcellulose 18.0g Talc powder 0.4g Lactose 0.2g Magnesium stearate 0.2g Anhydrous ethanol appropriate amount Make 100 pieces.

[0140] The specific preparation process is as follows: Take Carmaloidline G, ( S Ketamine, hydroxypropyl methylcellulose, talc, lactose, and magnesium stearate are mixed evenly, and anhydrous ethanol is added to make a soft mass. The mass is then passed through a 24-mesh sieve to form granules, dried, and magnesium stearate is added. The mixture is then mixed evenly and compressed into tablets.

[0141] Application Example 6 This invention discloses a capsule formulation using Carmaloidline G and fluoxetine as active pharmaceutical ingredients, the components of which are as follows: Carmaloidline G 3.5mg Fluoxetine 30.0mg 6.0g of starch Sodium metabisulfite 0.2g Magnesium stearate 0.2g Anhydrous ethanol appropriate amount Make 100 pills.

[0142] The specific preparation process is as follows: Carmaloidline G, fluoxetine, and sodium metabisulfite were mixed and then anhydrous ethanol was added to form a soft material. The material was passed through a 24-mesh sieve to form granules, dried, and then magnesium stearate was added. The mixture was then filled into capsules.

[0143] Application Example 7 This invention discloses an injectable formulation using Carmaloidline G and paroxetine as active pharmaceutical ingredients, the components of which are as follows: Carmaloidline G 5.0mg Paroxetine 20.0mg Vitamin C 0.2g Sodium chloride 6.0g Sodium bicarbonate 0.5mg 1000.0 mL of water for injection; Make 100 pieces.

[0144] The specific preparation process is as follows: After the above components are mixed, 100 vials can be obtained by using conventional injection preparation methods.

[0145] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. A compound and its pharmaceutically acceptable salt, solvate or crystal form, said compound being a cleaved C19 diterpenoid alkaloid having the structure shown in formula (1): （1）。 2. A pharmaceutical composition comprising the compound of claim 1 and its pharmaceutically acceptable salt, solvate or crystal form, and a pharmaceutically acceptable carrier, excipient or excipient.

3. The pharmaceutical composition according to claim 2, wherein the dosage form of the pharmaceutical composition is tablets, capsules, granules, oral liquids, powders, drops, inhalers, oral films, microcapsules, or injections.

4. The pharmaceutical composition according to claim 2, wherein the pharmaceutical composition is a liposome formulation, a nanoparticle formulation, a microemulsion formulation, or a solid dispersion formulation.

5. The use of the compound of claim 1 and its pharmaceutically acceptable salts, solvates or crystal forms in the preparation of an antidepressant.

6. The application according to claim 5, wherein the antidepressant is used to treat major depressive disorder, mild to moderate depressive disorder, or stress-related depressive disorder.

7. A method for preparing the compound of formula (1) in claim 1, comprising the following steps: The aconite was extracted with a solvent to obtain the extract, which was then concentrated to obtain the extract paste. The extract is dissolved in an acidic solution, and an acidic aqueous phase is obtained by extraction and separation. The acidic aqueous phase is then alkalized and extracted to obtain an organic phase. The organic phase was then subjected to column chromatography and high-performance liquid chromatography in sequence.

8. The preparation method according to claim 7, wherein the solvent is one or more selected from methanol, ethanol, acetone, chloroform, or petroleum ether; Optionally, the solvent is an aqueous ethanol solution with a volume fraction of 88% to 98%, the amount of solvent used is 8 to 10 times the mass of the plant raw material, and the extraction is carried out by reflux 2 to 4 times, each time for 1 to 3 hours.

9. The preparation method according to claim 7, wherein the acidic solution is dilute hydrochloric acid with a mass fraction of 0.1% to 1.0%; Optionally, the pH of the acidic aqueous phase after alkalization treatment is 8 to 11.

10. The preparation method according to any one of claims 7 to 9, wherein the high performance liquid chromatography mobile phase is an acetonitrile-water system with a volume ratio of (26 to 30):(70 to 74), and contains 0.01% to 0.5% trifluoroacetic acid; Optionally, the high-performance liquid chromatography uses a C18 column; Optionally, the column chromatography separation sequentially includes silica gel column chromatography, ODS column chromatography, and Sephadex LH-20 gel column chromatography; Optionally, the silica gel column chromatography uses a dichloromethane-methanol system for gradient elution; Optionally, the ODS column chromatography uses a methanol-water system as the mobile phase and contains 0.01% to 0.1% formic acid; Optionally, the Sephadex LH-20 gel column chromatography uses a methanol-water system as the mobile phase.

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