The use of mitoxanthone preparations in lymphatic tracing for use in thyroid-related surgeries.

Abstract

An application of a mitoxantrone preparation in preparation of a drug for diagnosing and treating a disease related to thyroidectomy, and use of mitoxantrone and / or a pharmaceutically acceptable salt thereof in preparation of a lymph tracer for preventing parathyroid gland staining in error, preventing parathyroid gland cutting in error, or reducing the rate of parathyroid gland cutting in error. Local injection of a mitoxantrone hydrochloride injection fluid would not cause parathyroid gland staining in error, and can reduce the rate of parathyroid gland cutting in error, thus protecting parathyroid glands well; moreover, no local or systemic toxic side effect is found after the local injection. The present invention has good tolerability, efficacy, and safety, and provides a new treatment idea in radically curing thyroid diseases thoroughly for patients suffering from diseases related to thyroidectomy.

Description

Use of a mitoxantrone preparation in the manufacture of a medicament for the diagnosis and treatment of a disease associated with thyroidectomy TECHNICAL FIELD

[0001] The present disclosure belongs to the field of pharmaceutical preparations, in particular to the use of mitoxantrone for lymphatic tracing in diseases associated with thyroidectomy. BACKGROUND

[0002] Thyroid cancer is the most common malignant tumor of the endocrine system and head and neck. The International Agency for Research on Cancer (IARC) reported in 2012 that the global incidence of thyroid cancer was 4.0 / 100,000, and the mortality rate was 0.5 / 100,000. The incidence of male was 1.9 / 100,000, and that of female was 6.1 / 100,000, with a male to female incidence ratio of 1:3. The mortality rate of male was 0.3 / 100,000, and that of female was 0.6 / 100,000, with a male to female mortality ratio of 1:2.

[0003] More than 90% of thyroid cancers are differentiated thyroid carcinomas (DTC). The main treatment methods for DTC include surgical treatment, postoperative I treatment and Thyroid Stimulating Hormone (TSH) suppression therapy. Among them, surgical treatment is the most important, directly affecting the follow-up treatment and follow-up of the disease, and is closely related to the prognosis. The thyroidectomy for DTC mainly includes total / near-total thyroidectomy and thyroid lobe+isthmus resection. Total thyroidectomy refers to the removal of all thyroid tissue, with no visible residual thyroid tissue; near-total thyroidectomy refers to the removal of almost all visible thyroid tissue (retaining <1g of non-tumor thyroid tissue, such as non-tumor thyroid tissue at the entrance of the recurrent laryngeal nerve into the larynx or at the parathyroid gland).

[0004] Although most DTC patients have a good prognosis, a low mortality rate, and a high 10-year survival rate, about 30% of DTC patients will relapse or metastasize, of which 2 / 3 occur within 10 years after surgery. Cervical lymph node metastasis is a risk factor for increased recurrence rate and decreased survival rate in DTC patients (especially those aged ≥45 years). 20% to 90% of DTC patients have cervical lymph node metastasis at the time of diagnosis, which often occurs in the central region of the neck. Therefore, the "Guidelines for the Diagnosis and Treatment of Thyroid Nodules and Differentiated Thyroid Carcinoma" suggests that prophylactic central lymph node dissection should be performed during DTC surgery under the condition of effectively preserving the parathyroid glands and recurrent laryngeal nerve.

[0005] Because the central lymph node dissection can easily injure the parathyroid glands, postoperative hypoparathyroidism is more common. Studies have shown that the incidence of permanent hypoparathyroidism after total and near-total thyroidectomy is 2%-33%. The reason is that the parathyroid glands are more likely to be mistakenly cut during surgery, which seriously affects the treatment effect of the surgery. Therefore, identifying and preserving as many parathyroid glands as possible becomes an important means to prevent postoperative hypoparathyroidism. Studies have found that the lymphatic network of the thyroid gland and the lymphatic network of the parathyroid gland are not connected, and the application of lymphatic tracers to the thyroid lymphatic drainage can well distinguish the thyroid gland from the parathyroid gland, avoiding the mistaken cutting of the parathyroid gland.

[0006] The currently recognized lymphatic tracing methods include dye method, nuclide method, and dye-nuclide combined tracing method.

[0007] The main dye-based lymph node tracers reported at home and abroad include methylene blue, nanocarbon, isothio blue, and patent blue. Currently, the most commonly used lymph node tracers in China are methylene blue and nanocarbon. Isothio blue and patent blue are less used in China due to their high price and difficulty in purchase.

[0008] Methylene blue, also known as methylene blue, is a water-soluble pigment dye with multiple clinical uses. It has been used as a lymph node tracer for many years, and its application in thyroid cancer has also accumulated certain clinical experience. Jozaghi et al. studied the application of methylene blue as a sentinel lymph node tracer in 300 cases of thyroid cancer and found that the lymph node detection rate was 68.8%, and the specificity was 100%, which proved that the application of methylene blue to help intraoperative tumor sentinel lymph node tracing can improve the intraoperative lymph node detection rate (Jozaghi Y, Richardson K, Anand S, et al. Frozen section analysis and sentinel lymph node biopsy in well differentiated thyroid cancer [J]. Journal of Otolaryngology-Head & Neck Surgery, 2013, 42(1): 1-5).

[0009] However, in the above-mentioned dye-based lymph node tracers, isosulfan blue, patent blue have weak protein binding force, and after injection, the tissue diffusion is less, the staining is fast, but the staining time is short, often requiring repeated injection, and the price is expensive, and there is no domestic manufacturer. Methylene blue can maintain the staining time for a long time, but has strong binding force with protein, and can also produce blue staining on the surrounding tissues. Nano-carbon has high lymphatic tropism, and accurate positioning, but it is not metabolized by the body, the production process is complex, and the staining speed is slow. The nuclide method has accurate positioning and simple intraoperative operation, but requires special detection instruments, has high cost, and due to the use of radioactive nuclides, there is a risk of nuclear pollution.

[0010] Therefore, it is an important means to improve the quality of life of patients with thyroid cancer and delay the life span of patients to develop a safe and effective lymph tracer for intraoperative lymph tracing of thyroid cancer, effectively locate the lymph nodes, predict whether the tumor has metastasis, and improve the lymph node cleaning rate while protecting the parathyroid glands.

[0011] SUMMARY

[0012] The present disclosure takes advantage of the characteristics of the lymphatic system tropism of mitoxantrone itself, which stains the lymph nodes near the thyroid cancer by its own color (deep blue), and develops it as a lymph tracer to stain the lymph nodes near the thyroid cancer in diseases related to thyroidectomy, such as thyroid cancer surgery, to help clinical lymph node positioning and cleaning and protect the parathyroid glands.

[0013] Therefore, the present disclosure aims to provide a use of mitoxantrone and / or a pharmaceutically acceptable salt thereof in the preparation of a lymph tracer for reducing parathyroid gland misstaining, reducing parathyroid gland misincision, or reducing the parathyroid gland misincision rate.

[0014] In the present disclosure, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by a person skilled in the art. And the protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, immunology related terms and laboratory operation steps used herein are the terms and conventional steps widely used in the corresponding field. At the same time, in order to better understand the present disclosure, the definitions and explanations of related terms are provided as follows.

[0015] It should also be understood that the terms used herein are only for the purpose of describing the specific embodiments and are not intended to be limiting.

[0016] As used herein, the terms "patient," "individual," and "subject" are used interchangeably and refer to any individual animal, more preferably a mammal (including, e.g., non-human animals such as cats, dogs, horses, rabbits, zoo animals, cows, pigs, sheep, and non-human primates) for which treatment is desired. In particular embodiments, the patient herein is a human. The patient can be suffering from, suspected of suffering from, or at risk of suffering from a thyroid tumor. As used herein, "disorder" is any condition that would benefit from treatment, including but not limited to chronic and acute disorders or diseases including those pathological conditions which predispose a mammal to the disorder in question.

[0017] As used herein, "pharmaceutical formulation" refers to a preparation which is in such form as to permit the biological activity of an active ingredient contained therein to be effective, and which contains no additional components which are unacceptably toxic to the subject to which the formulation would be administered.

[0018] As used herein, "pH adjusting agent" refers to a compound or mixture of compounds that can be used to ensure that the pH of a reconstituted kit is within the acceptable range for human or mammalian administration (pH about 4.0-10.5). Suitable pH adjusting agents include pharmaceutically acceptable buffers such as tricine, phosphate, or TRIS [i.e., tris(hydroxymethyl)aminomethane]; pharmaceutically acceptable acids such as pharmaceutically acceptable organic acids (e.g., formic acid, acetic acid) or mixtures thereof or inorganic acids (e.g., hydrochloric acid, phosphoric acid) or mixtures thereof, and pharmaceutically acceptable bases such as sodium carbonate, sodium bicarbonate, or mixtures thereof. When the conjugate used is in the acid salt form, the pH adjusting agent can optionally be provided in a separate vial or container so that the kit user can adjust the pH as part of a multi-step procedure.

[0019] As used herein, "pharmaceutically acceptable excipient" refers to an ingredient other than an active ingredient of a pharmaceutical formulation which is not toxic to a subject. Pharmaceutically acceptable excipients include, but are not limited to, buffers, carriers, stabilizers, or preservatives.

[0020] As used herein, "pharmaceutically acceptable salt" means a salt that is not biologically or otherwise undesirable. Pharmaceutically acceptable salts include acid and base addition salts. The phrase "pharmaceutically acceptable" means the material or composition must be chemically and / or toxicologically compatible with the other materials in the formulation and / or the mammal being treated therewith.

[0021] As used herein, "pharmaceutically acceptable acid addition salt" refers to those salts which are acceptable for use in pharmaceuticals and which are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and organic acids selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids such as formic, acetic, propionic, glycolic, gluconic, lactic, pyruvic, oxalic, malic, maleic, malonic, succinic, fumaric, tartaric, citric, aspartic, ascorbic, glutamic, anthranilic, benzoic, cinnamic, mandelic, embonic, phenylacetic, methanesulfonic, ethanesulfonic, p-toluenesulfonic and salicylic acids.

[0022] The term "pharmaceutically acceptable base addition salt" refers to those salts which are acceptable for use in pharmaceuticals and which are formed with organic or inorganic bases. Examples of acceptable inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, manganese and aluminum salts. Salts derived from pharmaceutically acceptable organic nontoxic bases include salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethylamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purines, piperizine, piperidine, N-ethylpiperidine and polyamine resins.

[0023] As used herein, "treatment" refers to clinical intervention in an attempt to alter the natural course of the individual being treated, and can be performed either for prophylia tic or in the full-fledged pathologic process. Desirable effects of treatment include, but are not limited to, preventing occurrence or reoccurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis.

[0024] As used herein, "administration" refers to the methods of giving a subject (e.g., a patient) a dose of a compound (e.g., mitoxantrone hydrochloride injection) or a pharmaceutical composition (e.g., a pharmaceutical composition comprising an inhibitor or antagonist). Administration can be performed by any suitable means, including parenterally, intrapulmonarily, and intranasally, and if desired for local treatment, intralesionally. Parenteral infusions include, for example, intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. Administration can be performed by any suitable route, for example, by injection, such as intravenous or subcutaneous injection, depending in part on whether the administration is short term or long term. Various dosing regimens are contemplated herein, including but not limited to single or multiple administrations at different points in time, bolus administration, and pulsed infusion.

[0025] As used herein, Full Analysis Set (FAS): refers to the data set of all subjects who participated in the trial, received treatment and had a baseline efficacy assessment according to the Intention To Treat (ITT) principle.

[0026] As used herein, Per Protocol Set (PPS): refers to the subset of the treated population who completed the trial and were excluded for serious protocol violations (i.e. study subjects who violated the inclusion criteria or exclusion criteria).

[0027] The present disclosure provides a use of mitoxantrone and / or a pharmaceutically acceptable salt thereof in the manufacture of a lymph tracer for reducing parathyroid gland mis-painting, reducing parathyroid gland mis-removal or reducing the rate of parathyroid gland mis-removal in a thyroidectomy-related disease.

[0028] The present disclosure also provides a method for reducing parathyroid gland mis-painting, reducing parathyroid gland mis-removal or reducing the rate of parathyroid gland mis-removal in a thyroidectomy-related disease, comprising administering mitoxantrone and / or a pharmaceutically acceptable salt thereof to a patient.

[0029] In one embodiment, the thyroidectomy-related disease is selected from a thyroid tumor or hyperthyroidism.

[0030] In one embodiment, the thyroid tumor comprises a thyroid benign tumor and a thyroid malignant tumor.

[0031] In one embodiment, the thyroid benign tumor is selected from a thyroid adenoma or a cyst.

[0032] In one embodiment, the thyroid malignant tumor is selected from a thyroid cancer or a thyroid malignant lymphoma.

[0033] In a preferred embodiment, the parathyroid gland mis-removal is parathyroid gland mis-removal in a thyroidectomy.

[0034] In a preferred embodiment, the thyroidectomy is selected from total thyroidectomy, near-total thyroidectomy, thyroid lobectomy with isthmectomy, partial thyroid lobectomy, thyroid adenoma removal, thyroid unilateral lobectomy and thyroid radical resection.

[0035] In a preferred embodiment, the lymph tracer is used for lymphatic mapping in thyroid cancer.

[0036] In a preferred embodiment, the lymph tracer comprises mitoxantrone and / or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0037] In a specific embodiment, the pharmaceutically acceptable excipient includes, but is not limited to, a buffer, a carrier, a stabilizer, or a preservative.

[0038] In a specific embodiment, the lymph tracer is an injection.

[0039] In a specific embodiment, the dosage form of the injection is a solution, a lyophilized powder, an emulsion, a liposome, a nanoparticle, a nanocrystal, a microcrystal, a microsphere, or a gel.

[0040] In a specific embodiment, the solution is sodium chloride injection or glucose injection.

[0041] In a preferred embodiment, the administration of the injection is selected from subcutaneous injection and intramuscular injection, preferably subcutaneous injection; preferably, the injection is local injection; preferably, the injection site is in the thyroid gland and / or the thyroid surrounding tissue organ; preferably, the injection volume of the injection with a concentration of 5 mg / ml is 0.2-1.2 ml.

[0042] In a preferred embodiment, the injection is multi-point injection, each injection point being spaced about 1 cm apart.

[0043] In a preferred embodiment, about 0.1 ml is injected at each point.

[0044] In a preferred embodiment, the total dose of both sides injection is not more than 0.6 ml.

[0045] In a preferred embodiment, the surgery is performed about 5 minutes after the injection.

[0046] In a specific embodiment, multi-point injection is performed on the thyroid gland, each injection point being spaced about 1 cm apart according to the size of the thyroid, about 0.1 ml is injected at each point, the total dose of both sides injection is not more than 0.6 ml, and the surgery is performed about 5 minutes later.

[0047] In a preferred embodiment, the pharmaceutically acceptable salt is those pharmaceutically acceptable salts of mitoxantrone formed with inorganic acids and organic acids.

[0048] In a specific embodiment, the inorganic acid is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, and phosphoric acid.

[0049] In one embodiment, the organic acid is selected from the group consisting of aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic organic acids, such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, pamoic acid, phenylacetic acid, methanesulfonic acid (mesylate), ethanesulfonic acid, p-toluenesulfonic acid and salicylic acid.

[0050] In one embodiment, the pharmaceutically acceptable salt is selected from the group consisting of mitoxantrone hydrochloride, mitoxantrone oxalate, mitoxantrone sulfate, mitoxantrone phosphate, mitoxantrone acetate and mitoxantrone citrate; more preferably, the pharmaceutically acceptable salt is mitoxantrone hydrochloride.

[0051] In a preferred embodiment, the lymphatic tracer comprises a pH adjusting agent.

[0052] In one embodiment, the pH adjusting agent is selected from one or more of hydrochloric acid, phosphoric acid, sulfuric acid, oxalic acid, acetic acid and citric acid.

[0053] In a preferred embodiment, the lymphatic tracer comprises an antioxidant.

[0054] In one embodiment, the antioxidant is selected from one or more of sodium sulfite, sodium bisulfite, sodium metabisulfite, sodium thiosulfate and disodium edetate; preferably sodium metabisulfite or disodium edetate.

[0055] In one embodiment, the lymphatic tracer comprises mitoxantrone or a salt thereof, sodium chloride, acetic acid, sodium acetate, sodium metabisulfite; more preferably, the lymphatic tracer further comprises sodium sulfate.

[0056] In one embodiment, the lymphatic tracer comprises mitoxantrone or a salt thereof, sodium chloride, acetic acid, sodium acetate, disodium edetate.

[0057] In one embodiment, the pH of the injection is in the range of 2.8-4.3.

[0058] In one embodiment, the amount of mitoxantrone or mitoxantrone in the salt thereof is in the range of 1-15 mg / ml; preferably 2-10 mg / ml; more preferably 2 mg / ml, 5 mg / ml or 10 mg / ml.

[0059] In one embodiment, the amount of sodium chloride is in the range of 3-18 mg / ml; preferably 4-16 mg / ml; more preferably 4 mg / ml, 8 mg / ml or 16 mg / ml.

[0060] In one embodiment, the content of the sodium chloride is 0.1-0.6 mg / ml; preferably 0.2-0.5 mg / ml; more preferably 0.2 mg / ml, 0.3 mg / ml or 0.4 mg / ml.

[0061] In one embodiment, the content of the sodium chloride is 0.1-0.6 mg / ml; preferably 0.2-0.5 mg / ml; more preferably 0.2 mg / ml, 0.3 mg / ml or 0.4 mg / ml.

[0062] In one embodiment, the content of the sodium chloride is 0.1-0.6 mg / ml; preferably 0.2-0.5 mg / ml; more preferably 0.2 mg / ml, 0.3 mg / ml or 0.4 mg / ml.

[0063] In one embodiment, the content of the sodium chloride is 0.1-0.6 mg / ml; preferably 0.2-0.5 mg / ml; more preferably 0.2 mg / ml, 0.3 mg / ml or 0.4 mg / ml.

[0064] In one preferred embodiment, the injection is prepared by the following method:

[0065] (1) Weigh the prescribed amount of sodium chloride, and mix it with solvent, dissolve, to obtain a solvent mixture; preferably, the solvent is water for injection; preferably, the solvent mixture is dissolved by stirring;

[0066] (2) Mix the solvent mixture obtained in step (1) with the prescribed amount of mitoxantrone and / or its pharmaceutically acceptable salt; preferably, it is dissolved by stirring; preferably, it is dissolved by stirring for 10-30 min.

[0067] In one embodiment, the method further comprises the following step: (3) filtration; preferably, filtration through 0.45 μm and / or 0.22 μm filter membrane.

[0068] In one embodiment, the method further comprises the following step: (4) nitrogen filling; preferably, after nitrogen filling, sterilization at 121 °C for 15 min.

[0069] In one embodiment, the pH value of the injection is in the range of 2.8-4.3.

[0070] In one preferred embodiment, the injection is prepared by the following method:

[0071] (1) Take the prescribed amount of acetic acid, sodium acetate, sodium chloride and edetate disodium; or take the prescribed amount of acetic acid, sodium acetate, sodium chloride, sodium pyrosulfite; or take the prescribed amount of acetic acid, sodium acetate, sodium chloride, sodium pyrosulfite, sodium sulfate, add the prescribed amount of water for injection, stir to dissolve;

[0072] (2) After dissolving, add the prescribed amount of mitoxantrone or its salt, stir for 10-30 min to dissolve;

[0073] (3) Filter through 0.45 μm and 0.22 μm filter membranes;

[0074] (4) Fill with nitrogen, 2 ml per bottle, cap, sterilize at 121°C for 15 min; the pH value is in the range of 2.8-4.3.

[0075] In one specific embodiment, the injection is prepared in a size of 2 ml: 10 mg.

[0076] The present disclosure can be used for the treatment of diseases related to thyroidectomy, such as thyroid cancer surgery, by locally injecting mitoxantrone hydrochloride injection, which can not only well stain the lymph nodes and accurately trace the lymph nodes, but also not cause false staining of the parathyroid glands, thereby reducing the parathyroid gland misincision rate, well protecting the parathyroid glands, and not producing local or systemic toxic side effects after local injection, and has good tolerability, effectiveness and safety, thereby providing a new treatment idea for patients with diseases related to thyroidectomy (such as thyroid cancer) in the thorough treatment of thyroid diseases. DETAILED DESCRIPTION

[0077] For the purpose of clarity and brevity, features are described herein as part of the same or separate embodiments, however, it will be understood that the scope of the disclosure can include embodiments having combinations of all or some of the described features.

[0078] Example 1 Preparation of Mitoxantrone Hydrochloride Injection Formula 1

[0079]

[0080] Take the prescribed amount of sodium chloride, acetic acid, sodium acetate and edetate disodium, add the prescribed amount of water for injection, stir to dissolve, add the prescribed amount of mitoxantrone hydrochloride after dissolving, stir for 30 min to dissolve, filter through 0.45 μm and 0.22 μm filter membranes, fill with nitrogen, cap, sterilize at 121°C for 15 min, and it is ready. The pH value is measured at 3.5.

[0081] Example 2 Preparation of Mitoxantrone Hydrochloride Injection Formula 2

[0082]

[0083] Weigh the prescribed amount of sodium chloride, acetic acid, sodium acetate, sodium pyrosulfite and sodium sulfate, add the prescribed amount of water for injection, stir to dissolve, after dissolving, add the prescribed amount of mitoxantrone hydrochloride, stir for 30 min to dissolve, filter through 0.45 μm and 0.22 μm filter membranes, fill and charge with nitrogen, roll on caps, sterilize at 121 °C for 15 min, and it is ready. The measured pH value is 3.4.

[0084] Preparation of Example 3 Mitoxantrone Hydrochloride Injection Formula 3

[0085]

[0086] Weigh the prescribed amount of sodium chloride, acetic acid, sodium acetate and disodium edetate, add the prescribed amount of water for injection, stir to dissolve, after dissolving, add the prescribed amount of mitoxantrone hydrochloride, stir for 30 min to dissolve, filter through 0.45 μm and 0.22 μm filter membranes, fill and charge with nitrogen, roll on caps, sterilize at 121 °C for 15 min, and it is ready. The measured pH value is 3.6.

[0087] Preparation of Example 4 Mitoxantrone Hydrochloride Injection Formula 4

[0088]

[0089] Weigh the prescribed amount of sodium chloride, acetic acid, sodium acetate, sodium pyrosulfite and sodium sulfate, add the prescribed amount of water for injection, stir to dissolve, after dissolving, add the prescribed amount of mitoxantrone hydrochloride, stir for 30 min to dissolve, filter through 0.45 μm and 0.22 μm filter membranes, fill and charge with nitrogen, roll on caps, sterilize at 121 °C for 15 min, and it is ready. The measured pH value is 3.7.

[0090] Preparation of Example 5 Mitoxantrone Hydrochloride Injection Formula 5

[0091]

[0092] Weigh the prescribed amount of sodium chloride, acetic acid, sodium acetate, sodium pyrosulfite and sodium sulfate, add the prescribed amount of water for injection, stir to dissolve, after dissolving, add the prescribed amount of mitoxantrone hydrochloride, stir for 30 min to dissolve, filter through 0.45 μm and 0.22 μm filter membranes, fill and charge with nitrogen, roll on caps, sterilize at 121 °C for 15 min, and it is ready. The measured pH value is 3.6.

[0093] Preparation of Example 6 Mitoxantrone Hydrochloride Injection Formula 6

[0094]

[0095]

[0096] Weigh the prescribed amount of sodium chloride, acetic acid, sodium acetate and edetate disodium, add the prescribed amount of water for injection, stir to dissolve, after dissolving, add the prescribed amount of mitoxantrone hydrochloride, stir for 30 min to dissolve, filter through 0.45 μm and 0.22 μm filter membranes, fill and nitrogenize, roll on caps, sterilize at 121 °C for 15 min, and it is ready. The pH value is determined to be 3.7.

[0097] Preparation of Example 7 Mitoxantrone Hydrochloride Injection Formula 7

[0098]

[0099] Weigh the prescribed amount of sodium chloride, acetic acid, sodium acetate and edetate disodium, add the prescribed amount of water for injection, stir to dissolve, after dissolving, add the prescribed amount of mitoxantrone hydrochloride, stir for 30 min to dissolve, filter through 0.45 μm and 0.22 μm filter membranes, fill and nitrogenize, roll on caps, sterilize at 121 °C for 15 min, and it is ready. The pH value is determined to be 3.7.

[0100] Preparation of Example 8 Mitoxantrone Hydrochloride Injection Formula 8

[0101]

[0102] Weigh the prescribed amount of sodium chloride, acetic acid, sodium acetate and edetate disodium, add the prescribed amount of water for injection, stir to dissolve, after dissolving, add the prescribed amount of mitoxantrone hydrochloride, stir for 30 min to dissolve, filter through 0.45 μm and 0.22 μm filter membranes, fill and nitrogenize, roll on caps, sterilize at 121 °C for 15 min, and it is ready. The pH value is determined to be 3.7.

[0103] Example 9 Pharmacokinetics and Pharmacodynamics Study of Mitoxantrone Hydrochloride Injection

[0104] The organ target of mitoxantrone hydrochloride injection for lymph tracing is the thyroid drainage area lymph node. When mitoxantrone hydrochloride is combined with hydrochloric acid, a uniform acidic solution is formed. After the injection is administered in the tissue space, the pH of the microenvironment changes, and mitoxantrone hydrochloride gradually precipitates nanocrystals. The crystals prevent it from entering the blood circulation through capillaries. Due to the strong permeability of capillary lymphatic vessels, it can enter capillary lymphatic vessels through the intercellular space and pinocytosis and phagocytosis of endothelial cells, then pass through the lymphatic drainage to the regional lymph nodes, and accumulate in the lymph nodes for a period of time, achieving the effect of lymph node staining and tracing.

[0105] In order to investigate the safety and effectiveness of mitoxantrone hydrochloride injection for lymphatic tracing of cancer drainage lymph nodes in patients with thyroid cancer, and the tolerability and pharmacokinetics of mitoxantrone hydrochloride injection in thyroid cancer subjects, and determine the safe dose range, the present example adopts a single-center, randomized, open, blank control trial design. After sufficient exposure of the thyroid gland, multiple injections of mitoxantrone hydrochloride injection are made on the thyroid gland. The total injection dose does not exceed 0.6 ml according to the size of the thyroid gland. The tolerability and pharmacokinetics of the subjects are studied in groups, and the effectiveness of the test drug is observed at the same time.

[0106] Pharmacokinetic results: After peritumoral injection of mitoxantrone hydrochloride injection, rapid absorption was observed, with peak concentration at 10 min after injection in most cases. The drug was rapidly eliminated after entering the blood, and almost completely eliminated from the plasma 30 min after administration. In addition, the overall blood concentration after administration showed a dose-dependent trend. The blood concentrations of different subjects at each time point in the low-dose group were all below the lower limit of quantification. In the medium and high-dose groups, the blood concentrations of most patients at most time points were above the lower limit of quantification, with the highest concentration detected being only 13.10 ng / ml. In the prior art document (Zhu Jianming et al., "Parathyroid gland injury and postoperative hypocalcemia in thyroid surgery", China Modern Surgery Journal, April 2010, Vol. 14, No. 2, pp. 112-114), high-dose mitoxantrone chemotherapy was used to treat ovarian cancer. The maximum tolerated total dose of mitoxantrone was 75 mg / m 2 , and the AUC at this dose was 560-1700 ng·h / ml, which was 135.4-411.0 times the maximum AUC (248.15 ng·min / ml) in this trial. Therefore, peritumoral injection of mitoxantrone hydrochloride injection does not cause toxic side effects and does not pose a safety risk after local injection.

[0107] Pharmacodynamic results: None of the subjects showed discoloration of the stained lymph nodes after injection of the tracer until the end of lymph node dissection, so the success rate of tracer persistence was 100%.

[0108] Example 10 Application of mitoxantrone hydrochloride injection in lymphatic tracing of thyroid cancer surgery patients

[0109] 1. Clinical trial design

[0110] In this example, a multi-center, randomized clinical trial design is adopted. Subjects who meet the requirements of the scheme are randomly allocated in a 1:1 ratio to compare the effectiveness and safety of mitoxantrone hydrochloride injection in the treatment of thyroid cancer patients. The specific requirements of the scheme are as follows:

[0111] • Age 18-70 years (including 18 and 70 years), male and female

[0112] • Preoperative clinical diagnosis of thyroid cancer, planning to undergo radical operation for thyroid cancer

[0113] • According to preoperative assessment, planning to undergo total thyroidectomy plus total central lymph node dissection

[0114] • No obvious surgical contraindication was found in routine preoperative examination

[0115] • Can communicate well with the researchers and can complete the study according to the study

[0116] • The patient voluntarily participates in this study and signs the informed consent form

[0117] The test group (administering mitoxantrone hydrochloride injection, the content of the effective component in mitoxantrone hydrochloride injection is 5mg / ml, multi-point injection on the thyroid body, according to the size of the thyroid, each injection point is about 1cm apart, each point is about 0.1ml, the total dose of both sides is not more than 0.6ml, about 5min for operation) and the control group (patients with conventional surgery, without the use of mitoxantrone hydrochloride injection) were examined and recorded the number of parathyroid glands removed, the total number of lymph node removed, and the number of metastatic lymph nodes.

[0118] Safety evaluation index:

[0119] Safety evaluation will be throughout the study. Subjects who withdraw early will be evaluated for safety before withdrawal. Safety evaluation includes changes in vital signs, physical examination, electrocardiogram, clinical laboratory test indicators (blood routine, urine routine, blood biochemistry, coagulation function, pregnancy test, etc.), adverse events and serious adverse events, and early withdrawal due to safety or tolerability reasons.

[0120] 2. Clinical trial effectiveness results

[0121] The results of the clinical trial showed that the parathyroid gland misincision rate of conventional surgery was about 27%, and the parathyroid gland misincision rate using the product of the present disclosure was less than 7%, and the parathyroid gland misincision rate and the number of parathyroid gland misincision of individuals using the product of the present disclosure were significantly lower than those of conventional surgery. In this study, the evaluation of the main efficacy indicators was carried out using PPS data set and FAS data set for statistical analysis, and the conclusions were consistent. And the results of sensitivity analysis were also consistent. There was a significant difference in the main efficacy indicators between the test group and the control group (P<0.001), and no obvious adverse reactions were found.

[0122] 3. Safety evaluation results of clinical trial

[0123] Phase I clinical trials showed that a very small amount of mitoxantrone hydrochloride injection for lymph tracing entered the blood circulation after local injection, and did not produce systemic toxic side effects, with good safety. Phase II and III clinical trials showed that when using mitoxantrone hydrochloride injection for lymph tracing for thyroid cancer, the percentage of adverse events (AEs), the percentage of serious adverse events (SAEs), and the percentage of AEs above grade 3 were similar in the test group and the blank control group, and no AEs and SAEs related to the test drug occurred. Using chi-square test for inter-group comparison, the differences in the incidence of AEs, SAEs, and AEs above grade 3 between the two groups of subjects were not statistically significant. Therefore, it can be concluded that the safety results of the test group and the blank control group are similar and good.

[0124] The results of the above clinical trial showed that:

[0125] (1) Mitoxantrone hydrochloride injection has good parathyroid gland protection effect;

[0126] (2) Mitoxantrone hydrochloride injection has good lymph node staining performance and can accurately trace lymph nodes;

[0127] (3) The test drug does not cause false staining of the parathyroid glands;

[0128] (4) The use of the test drug in thyroid surgery can remove more lymph nodes;

[0129] (5) The test group and the control group had significant differences in the number of lymph nodes with sizes of ≤2 mm, >2 mm, ≤5 mm, and >5 mm, ≤10 mm, but no significant difference was found in the number of lymph nodes with sizes of >10 mm;

[0130] (6) No local or systemic toxic side effects were observed after local injection of the test drug, and no adverse events and adverse reactions related to the test drug were observed, indicating good tolerance and safety.

[0131] From the above results, it can be concluded that the clinical trial has good effectiveness and safety, and the use of mitoxantrone hydrochloride injection for lymph node staining can accurately locate and remove more lymph nodes, reduce parathyroid gland removal, and reduce the parathyroid gland removal rate, providing a new treatment approach for the complete radical treatment of thyroid cancer in patients with thyroid cancer.

[0132] Example 11 Application of Mitoxantrone Hydrochloride Injection in Lymph Tracing in Patients Undergoing Radical Thyroidectomy for Thyroid Cancer

[0133] Preoperative diagnosis: thyroid cancer

[0134] Postoperative diagnosis: thyroid cancer

[0135] Operation name: Radical resection of thyroid carcinoma (bilateral total thyroidectomy + bilateral VI, VII group lymph node dissection)

[0136] Anesthesia: General anesthesia

[0137] Operation steps: After checking the patient and successful anesthesia, the patient's shoulder is padded with a pillow, the neck is tilted back, and the iodine complex is sterilized and laid on the surgical field. An arc-shaped incision about 7 cm long is made at the lower part of the neck where the skin lines coincide. The skin, subcutaneous tissue, platysma, and anterior cervical fascia are cut layer by layer. The skin flap is separated upward and downward between the platysma and the anterior cervical fascia. The anterior cervical fascia is cut on the midline, and the anterior cervical muscle is separated to the surface of the thyroid gland. Exploration shows that the thyroid gland is soft, with multiple nodules on both lobes. The right lobe has a hard nodule about 1 cm in size. At 1 cm intervals above and below the left lobe of the thyroid gland and 1 cm intervals above and below the right lobe, 0.1 ml of lymph tracer mitoxantrone hydrochloride injection (total 0.4 ml) is injected under the capsule of the thyroid gland. The drug is recovered after 5 minutes of pressure on the puncture site. The thyroid gland and lymph nodes gradually turn blue. Right thyroidectomy is performed first. The right lobe of the thyroid gland is separated laterally, the right middle thyroid vein is disconnected, the prelaryngeal lymph node and thyroid pyramidal lobe are removed, the isthmus is disconnected near the right side, the right lobe of the thyroid gland is pulled up to the left side, the upper and lower polar vessels of the right lobe are disconnected, the recurrent laryngeal nerve is carefully separated on the dorsal side of the gland, and the right lobe of the thyroid gland is completely removed. The left lobe is removed in a similar manner. The bilateral recurrent laryngeal nerves are completely separated, and the VI and VII group lymphatic adipose tissue is removed under the guidance of the lymph tracer (up to the hyoid bone level, lateral to the carotid artery to the medial margin, and down to the superior margin of the innominate artery). During the process, the visible parathyroid glands are preserved in situ. The wound is carefully stopped bleeding, and the gauze and instruments are counted. Absorbable sutures are used to close the anterior cervical fascia and subcutaneous tissue layer by layer, and the wound is adhered. The operation is successful with minimal bleeding and no blood transfusion. The patient is successfully extubated after the operation, has no hoarseness, and returns safely to the ward. The removed tissue is sent for pathological examination.

[0138] Pathological examination results: Metastatic carcinoma of lymph nodes (0 / 0 in the right paratracheal region; 1 / 1 in the pretracheal region; 1 / 6 in the left paratracheal region; 0 / 0 in the prelaryngeal region), with a small amount of thyroid tissue visible in the right paratracheal and pretracheal regions.

[0139] No parathyroid gland was mistakenly removed. A total of 7 lymph nodes were found, of which 2 were ≤2 mm in size, 5 were >2 mm and ≤5 mm in size, and 0 were >5 mm and ≤10 mm in size. The above-mentioned lymph nodes were dyed, with a dyeing rate of 100%.

Claims

DEPCT6616 / 02 / 25661. The use of mitoxanthone and / or its pharmaceutically acceptable salts in serological tracing preparations to prevent parathyroid staining errors, prevent parathyroidectomy errors, or reduce the parathyroidectomy error rate for thyroidectomy-related diseases. Ideally, thyroidectomy-related diseases should be selected from thyroid tumors or hyperthyroidism; ideally, thyroid tumors should include both benign and malignant thyroid tumors; ideally, benign thyroid tumors should be selected from thyroid tumors or cysts.Ideally, malignant thyroid tumors should be selected from thyroid epithelial cancer or malignant thyroid lymphoma.

2. Methods for preventing parathyroid staining errors, preventing parathyroidectomy errors, or reducing the parathyroidectomy error rate for thyroidectomy-related diseases include: administering mitoxanthone and / or pharmaceutically acceptable salts to the patient; ideally, thyroidectomy-related diseases should be selected from thyroid tumors or hyperthyroidism; ideally, thyroid tumors should include both benign and malignant thyroid tumors; ideally, benign thyroid tumors should be selected from thyroid tumors or cysts;Ideally, malignant thyroid tumors should be selected from thyroid epithelial cancer or malignant thyroid lymphoma.

3. Use under claim 1 or method under claim 2 where parathyroidectomy error is the failure to remove the parathyroid glands during thyroidectomy.

4. Use or method under the preceding claim where thyroidectomy is selected from total thyroidectomy, near-total thyroidectomy, lobe resection + midsection for thyroid, total thyroidectomy, unilateral lobe resection for thyroid, and thyroidectomy with cervical lymph nodes; and ideally, a lymphoid tracer should be used to trace the lymphoid system in thyroid epithelial cancer.

5. Use or method under the preceding claim where the lymphoid tracer contains mitoxanthone and / or pharmaceutically acceptable salts of such substance and a pharmaceutically acceptable stabilizer;Ideally, pharmaceutically acceptable drug stabilizers should include, but are not limited to, the following: buffers, carriers, stabilizers, or preservatives; ideally, the serological tracer should be an injectable substance; ideally, the injectable substance should be in the form of a solution, a low-temperature solidified powder, an emulsion, a liposome, a nanoparticle, a nanocrystal, a microcrystal, a microsphere, or a gel; and ideally, the injectable substance in solution form should be a sodium chloride or glucose solution.

6. Use or method as per the prior claims. The injected substance will be administered subcutaneously or intramuscularly, and ideally subcutaneously; ideally, the substance should be delivered locally; ideally, the injection site should be on the thyroid gland and / or surrounding tissues and organs; ideally, 0.2-1.2 mL of substance at a concentration of 5 mg / mL should be administered; ideally, the substance should be administered at multiple points spaced approximately 1 cm apart; ideally, approximately 0.1 mL of substance should be administered at each point; ideally, the total amount of substance administered on both sides should not exceed 0.6 mL.And ideally, the surgery should be performed approximately 5 minutes after the injection.

7. The use or method according to the prior claims where the pharmaceutically acceptable salt is a pharmaceutically acceptable salt constructed from mitoxanthone and inorganic or organic acids; ideally, the inorganic acid should be selected from hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, and phosphoric acid; ideally, the organic acid should be selected from aliphatic, cycloaliphatic, aromatic, and... Raliphatic, heterocyclic, carboxylic, and sulfonic organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, pamoic acid, phenylacetic acid, methanesulfonic acid (methanesulfonate), ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid;Ideally, the pharmaceutically acceptable salt should be selected from mitoxantrone hydrochloride, mitoxantrone oxalate, mitoxantrone sulfate, mitoxantrone phosphate, mitoxantrone acetate, and mitoxantrone citrate, and even better, the pharmaceutically acceptable salt should be mitoxantrone hydrochloride.

8. The use or method according to the preceding claims where the serological tracer has a pH regulator; and ideally, the pH regulator should be one or more of the following substances, which are selected from: The group consisting of hydrochloric acid, phosphoric acid, sulfuric acid, oxalic acid, acetic acid, and citric acid.

9. The use or method according to the prior claims where the lymphatic tracer contains an antioxidant; ideally, the antioxidant should be one or more of the substances chosen from the group consisting of sodium sulfite, sodium bisulfite, sodium pyrosulfite, sodium thiosulfate and disodium editate, and ideally the antioxidant should be sodium pyrosulfite or disodium editate;Ideally, the lymphatic tracer should contain mitoxantone or its salts, sodium chloride, acetic acid, sodium acetate, and sodium pyrosulfite; and even better, it should also contain sodium sulfate. Ideally, the lymphatic tracer should contain mitoxantone or its salts, sodium chloride, acetic acid, sodium acetate, and disodium edetate; ideally, the pH of the injectable solution should be in the range of 2.8-4.3; ideally, the amount of mitoxantone or mitoxantone in the salt... The concentration of the aforementioned substances should be 1-15 mg / mL, ideally 2-10 mg / mL, and even better, 2 mg / mL, 5 mg / mL, or 10 mg / mL in terms of weight by volume; the sodium chloride concentration should be 3-18 mg / mL, ideally 4-16 mg / mL, and even better, 4 mg / mL, 8 mg / mL, or 16 mg / mL in terms of weight by volume; the acetic acid concentration should be 0.15-1 mg / mL, ideally 0.23-0.92 mg / mL, and even better, 0.23 mg / mL, 0.46 mg / mL, or 0.92 mg / mL in terms of weight by volume;Ideally, the sodium acetate content should be 0.03-0.15 mg / mL, ideally 0.05-0.1 mg / mL, and even better, 0.05 mg / mL or 0.1 mg / mL in terms of weight by volume; ideally, the antioxidant content should be 0.05-0.5 mg / mL, ideally 0.08-0.4 mg / mL, and even better, 0.1 mg / mL, 0.2 mg / mL, or 0.3 mg / mL in terms of weight by volume; and ideally, the sodium sulfate content should be 0.05-0.6 mg / mL, ideally 0.15-0.45 mg / mL, and even better... It should be 0.15mg / mL, 0.3mg / mL or 0.45mg / mL in terms of weight by volume.

10. Use or method according to the preceding claims where the injectable substance shall be prepared by the following methods:(1) weighing acetic acid, sodium acetate, sodium chloride and disodium editate in predetermined amounts or weighing acetic acid, sodium acetate, sodium chloride and sodium pyrosulfite in predetermined amounts or weighing acetic acid, sodium acetate, sodium chloride;(1) Mix the predetermined amounts of sodium pyrosulfite and sodium sulfate with the solvent and dissolve them to obtain a mixed solution of the drug stabilizer. Ideally, the solvent should be aqueous for injection, and ideally, the drug stabilizer should be dissolved by stirring; and (2) Mix the mixed solution of the drug stabilizer obtained from step (1) with mitoxantrone and / or pharmaceutically acceptable salts of such substance in predetermined amounts. Ideally, the mitoxantrone and / or pharmaceutically acceptable salts of such substance should be dissolved by stirring. Dissolution by stirring: Ideally, the mitoxanthone and / or pharmaceutically acceptable salts of such substance should be dissolved by stirring for 10–30 minutes; ideally, the method should also include the following steps:(3) Filtration: Ideally, the filtration should be performed using a thin membrane of 0.45 µm and / or 0.22 µm; ideally, the method should also include the following steps:(4) Bottling and nitrogen gas filling: Ideally, sterilization should be performed at 121 °C for 15 minutes after nitrogen gas filling;Ideally, the pH of the injectable substance should be in the range of 2.8-4.3; and ideally, the injectable substance should be prepared according to specific specifications with a ratio of 2mL:10mg;