Ointment of rifamycin-quinolidinone coupling molecule and its preparation method

Abstract

A rifamycin-quinolizinon conjugate molecule ointment is provided. The raw material components of the rifamycin-quinolizinon conjugate molecule ointment, based on a total mass percentage of 100%, include 0.05% to 0.7% of the rifamycin-quinolizinon conjugate molecule, 55% to 65% of polyethylene glycol 400, 25% to 35% of polyethylene glycol 3350, 8% to 12% of propylene glycol, 0.1% to 1% of a solubilizer, and 0.3% to 1% of an antioxidant. The rifamycin-quinolizinon conjugate molecule ointment can improve the pharmaceutical content and stability of the pharmaceutical components of the formulation, thereby improving dosing efficacy and user experience. [Formula 1] JPEG2025525986000016.jpg54158

Description

[Technical Field]

[0001] The present application relates to an ointment of rifamycin-quinolidinone coupling molecule and its preparation method, which belongs to the technical field of medicine. [Background technology]

[0002] The rifamycin-quinolidinone coupling molecule is a multi-target drug, the structural formula of which is shown as Formula I below. [ka]

[0003] This molecule consists of a rifamycin and a quinolone pharmacophore and can effectively treat bacterial infections. The rifamycin-quinolidinone coupling molecule exhibits low water solubility, being soluble in dichloromethane, polyethylene glycol 400, and dimethyl sulfoxide, and practically insoluble in ethanol, methanol, and diethyl ether. The water solubility of this molecule is pH-dependent, with maximum solubility observed at pH 10 and relatively low solubility at lower pH values. Furthermore, the rifamycin-quinolidinone coupling molecule is easily oxidized upon exposure to air, converting from its phenolic form to its quinone form. At temperatures above 60°C, this molecule can degrade. Under dry storage conditions, the primary degradation product is its quinone form. Oxidation or decomposition of the compound can result in a decrease in the efficacy of the effective dose of the drug. Therefore, a series of improvements or developments in specific formulations are needed to broaden its application prospects. Summary of the Invention

[0004] In view of the above-mentioned shortcomings in the prior art, the objective of the present application is to provide an ointment of rifamycin-quinolidinone coupling molecule, which can improve the drug loading capacity of the formulation and the stability of the drug components, prevent the formation of oxidation products, improve the medication effect, and enhance the medication safety.

[0005] The present application also aims to provide a method for preparing an ointment of a rifamycin-quinolidinone coupling molecule, which can improve drug loading and product stability.

[0006] The objectives of this application are achieved by the following technical solutions: In one aspect, the present application provides a rifamycin-quinolidinone coupled molecule ointment, wherein the components of the rifamycin-quinolidinone coupled molecule ointment include a rifamycin-quinolidinone coupled molecule shown in Formula I, polyethylene glycol 400, polyethylene glycol 3350, propylene glycol, a solubilizer, and an antioxidant; [ka] Based on a total mass percentage of 100%, the content of the rifamycin-quinolidinone coupling molecule is 0.05% by mass to 0.7% by mass, the content of polyethylene glycol 400 is 55% by mass to 65% by mass, the content of polyethylene glycol 3350 is 25% by mass to 35% by mass, the content of propylene glycol is 8% by mass to 12% by mass, the content of the solubilizer is 0.1% by mass to 1% by mass, and the content of the antioxidant is 0.3% by mass to 1% by mass.

[0007] In some embodiments, the content of the rifamycin-quinolidinone coupled molecule is 0.3% to 0.7% by mass.

[0008] In some embodiments, the content of the rifamycin-quinolidinone coupled molecule is 0.05% to 0.5% by mass.

[0009] In some embodiments, the content of the rifamycin-quinolidinone coupled molecule is 0.5% by weight.

[0010] In some embodiments, the content of polyethylene glycol 400 is 55% to 60% by mass.

[0011] In some embodiments, the content of polyethylene glycol 400 is 56% to 58% by mass.

[0012] In some embodiments, the content of polyethylene glycol 3350 is 28% to 33% by mass.

[0013] In some embodiments, the content of polyethylene glycol 3350 is 30% to 32% by mass.

[0014] In some embodiments, the solubilizer is present in an amount of 0.3% to 0.7% by weight.

[0015] In some embodiments, the solubilizing agent comprises triethanolamine.

[0016] In some embodiments, the antioxidant content is 0.5% to 1% by weight.

[0017] In some embodiments, the antioxidant comprises one or more of vitamin C and vitamin E.

[0018] In some embodiments, the content of vitamin C and / or vitamin E is 0.3% to 0.5% by weight.

[0019] In some embodiments, the components of a rifamycin-quinolidinone coupled molecule ointment include a rifamycin-quinolidinone coupled molecule of Formula I, polyethylene glycol 400, polyethylene glycol 3350, propylene glycol, a solubilizer, and an antioxidant; based on a total mass percentage of 100%, the content of the rifamycin-quinolidinone coupled molecule is 0.3% to 0.7% by mass, the content of polyethylene glycol 400 is 55% to 60% by mass, the content of polyethylene glycol 3350 is 28% to 33% by mass, the content of propylene glycol is 8% to 12% by mass, the content of the solubilizer is 0.1% to 1% by mass, and the content of the antioxidant is 0.5% to 1% by mass. In the above rifamycin-quinolidinone coupled molecule ointment, the solubilizer preferably includes triethanolamine. In the above ointment of rifamycin-quinolidinone coupling molecule, the antioxidant preferably includes one or more of vitamin C and vitamin E.

[0020] In some embodiments, the components of the rifamycin-quinolidinone coupled molecule ointment, based on a total mass percentage of 100%, include 0.5% by weight of the rifamycin-quinolidinone coupled molecule, 56% to 58% by weight of polyethylene glycol 400, 30% to 32% by weight of polyethylene glycol 3350, 10% by weight of propylene glycol, 0.3% to 0.7% by weight of triethanolamine, 0.3% to 0.5% by weight of vitamin C, and 0.3% to 0.5% by weight of vitamin E.

[0021] In another aspect, the present application provides a method for preparing a rifamycin-quinolidinone coupled molecule ointment described herein, comprising: heating, melting, and mixing polyethylene glycol 3350, propylene glycol, a solubilizing agent, an antioxidant, and a portion of polyethylene glycol 400 to obtain a first mixture; mixing and dissolving the rifamycin-quinolidinone coupled molecule and the remainder of the polyethylene glycol 400 to obtain a first medicinal solution; and mixing the first medicinal solution with the first mixture to obtain the rifamycin-quinolidinone coupled molecule ointment.

[0022] In some embodiments, in the step of mixing and dissolving the rifamycin-quinolidinone coupling molecule and the remainder of the polyethylene glycol 400 to obtain the first drug solution, the mixing and dissolving is performed at a temperature of 50°C.

[0023] In some embodiments, in the step of mixing the first chemical liquid with the first mixture, the mixing is performed at a temperature of 50 to 53°C.

[0024] In another aspect, the present application provides a method for preparing a rifamycin-quinolidinone coupled molecule ointment described herein, comprising: heating, melting, and mixing polyethylene glycol 3350, propylene glycol, a solubilizing agent, an antioxidant, and polyethylene glycol 400 to obtain a first mixture; and mixing the rifamycin-quinolidinone coupled molecule with the first mixture to obtain the rifamycin-quinolidinone coupled molecule ointment.

[0025] In some embodiments, the heat melting is carried out at a temperature of 70-73°C.

[0026] In some embodiments, in the step of combining the rifamycin-quinolidinone coupled molecule with the first mixture, the combining is performed at a temperature of 50°C. Specific features of the invention to which this application pertains are set forth in the appended claims. The features and advantages of the invention to which this application pertains may be more fully understood with reference to the exemplary embodiments and drawings described in detail below, which are briefly described as follows: [Brief explanation of the drawings]

[0027] [Figure 1] 1 shows the oxidative decomposition of the rifamycin-quinolidinone coupled molecule, resulting in the production of oxidized impurities. [Figure 2] 1 shows comparative electron microscopy results of formulations with different solubilizers. [Figure 3] 1 shows the bacterial load results in mice from Example 5 of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0028] The technical solutions of the present disclosure are described in detail below to provide a clearer understanding of the technical features, objectives, and beneficial effects of the present disclosure. However, this description should not be construed as limiting the embodiments of the present disclosure. In the following examples, unless otherwise specified, all experimental methods are conventional methods; reagents and materials are commercially available unless otherwise specified.

[0029] Definition of Terms In this application, the term "ointment" generally refers to a homogeneous semi-solid topical formulation obtained by mixing a drug with an oily or water-soluble base. Ointments can be classified into solution-type ointments and suspension-type ointments due to differences in the dispersion state of the drug in the base. Solution-type ointments are prepared by dissolving or co-dissolving a drug in a base or base components, while suspension-type ointments are prepared by uniformly dispersing a drug fine powder in a base. The ointments provided by the present application are generally used for topical and local treatments, and in certain embodiments, the drug in the ointment (e.g., a compound of Formula I described herein) can exert a systemic pharmacological or therapeutic effect upon transdermal absorption. In certain embodiments, the base of an ointment formulation serves as an excipient and pharmaceutical carrier, which can have a significant impact on the quality of the ointment and the release and absorption of the drug. Ointment formulation bases commonly used in the art can be classified into oily bases, emulsion bases, and water-soluble bases. The ointment formulations required for the treatment methods of the present application may be obtained using suitable preparation methods and processes in the art.

[0030] In this application, the term "melting" generally refers to a first-order phase transition process accompanied by an increase in enthalpy, entropy, and volume. For example, as the temperature increases, the kinetic energy of the thermal motion of molecules increases, resulting in the destruction of crystals. The transition process of a substance from a crystalline phase to a liquid phase can be referred to as melting. For example, in this application, the process of heating a substance that is solid at room temperature to a certain temperature to reach its melting point and become a liquid substance, thereby acquiring the physical properties of a liquid, can also be referred to as melting. A substance in a molten state can be in a liquid state or in a coexisting solid-liquid state.

[0031] In this application, the term "mixing" generally refers to the process of combining one or more compounds, cells, molecules, etc. in the same area. This process can be carried out, for example, in a test tube, a culture dish, or any container that allows for the mixing of one or more compounds, cells, or molecules.

[0032] In this application, all numbers disclosed herein are approximate, regardless of whether the word "about" or "approximately" is used, and the numerical value of each number can vary, for example, by ±1%, ±2%, ±5%, etc. The term "about" or "approximately" generally refers to an acceptable error range for a particular value as determined by one of ordinary skill in the art, which may depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, "about" or "approximately" can mean within one standard deviation or more than one standard deviation, in accordance with the practice in the art. Alternatively, "about" or "approximately" can mean a range of up to 10% or 20% (i.e., ±10% or ±20%). For example, about 3 mg can include any number between 2.7 mg and 3.3 mg (for 10%) or between 2.4 mg and 3.6 mg (for 20%). Furthermore, particularly in the context of biological systems or processes, the term can mean a value up to an order of magnitude or 5-fold. Unless otherwise specified, when a particular value or composition is provided in this specification and the appended claims, the meaning of "about" or "approximately" should be assumed to be within an acceptable range of error for the particular value or composition.

[0033] Detailed Description of the Invention In one aspect, the present application provides a rifamycin-quinolidinone coupled molecule ointment, the components of which include the rifamycin-quinolidinone coupled molecule compound of Formula I, polyethylene glycol 400, polyethylene glycol 3350, propylene glycol, a solubilizer, and an antioxidant; based on a total mass percentage of 100%, the content of the rifamycin-quinolidinone coupled molecule is 0.05% by mass to 0.7% by mass, the content of polyethylene glycol 400 is 55% by mass to 65% by mass, the content of polyethylene glycol 3350 is 25% by mass to 35% by mass, the content of propylene glycol is 8% by mass to 12% by mass, the content of the solubilizer is 0.1% by mass to 1% by mass, and the content of the antioxidant is 0.3% by mass to 1% by mass.

[0034] In another aspect, the present application provides a method for preparing a rifamycin-quinolidinone coupled molecule ointment described herein, comprising: heating, melting, and mixing polyethylene glycol 3350, propylene glycol, a solubilizing agent, an antioxidant, and a portion of polyethylene glycol 400 to obtain a first mixture; mixing and dissolving the rifamycin-quinolidinone coupled molecule and the remainder of the polyethylene glycol 400 to obtain a first medicinal solution; and mixing the first medicinal solution with the first mixture to obtain the rifamycin-quinolidinone coupled molecule ointment.

[0035] In another aspect, the present application provides a method for preparing a rifamycin-quinolidinone coupled molecule ointment described herein, comprising: heating, melting, and mixing polyethylene glycol 3350, propylene glycol, a solubilizing agent, an antioxidant, and polyethylene glycol 400 to obtain a first mixture; and mixing the rifamycin-quinolidinone coupled molecule with the first mixture to obtain the rifamycin-quinolidinone coupled molecule ointment.

[0036] Rifamycin-quinolidinone coupling molecule The rifamycin-quinolidinone coupled molecule (API) is a multi-target compound, which consists of a rifamycin and a quinolone pharmacophore and has the structural formula shown as Formula I, [ka] This molecule has antibacterial activity against bacteria such as Staphylococcus, Streptococcus, and Streptococcus pneumoniae, and its topical formulation can be used to treat skin infections. The drug acts on three different targets, including RNA polymerase, DNA gyrase, and topoisomerase IV. This multitarget mechanism of action can achieve the technical effect of reducing the possibility of drug resistance. The rifamycin-quinolidinone coupling molecule is prepared by TenNor Therapeutics.

[0037] In some embodiments, the amount of the rifamycin-quinolidinone coupled molecule is 0.05% to 0.7% by weight, for example, about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.1%, about 0.15%, about 0.2%, about 0.25%, about 0.3%, about 0.35%, about 0.4%, about 0.45%, about 0.5%, about 0.55%, about 0.6%, about 0.65%, or about 0.7% of the total weight.

[0038] In some embodiments, the amount of the rifamycin-quinolidinone coupled molecule is 0.3% to 0.7% by weight, e.g., about 0.3%, about 0.35%, about 0.4%, about 0.45%, about 0.5%, about 0.55%, about 0.6%, about 0.65%, or about 0.7% of the total weight.

[0039] In some embodiments, the amount of the rifamycin-quinolidinone coupled molecule is 0.05% to 0.5% by weight, e.g., about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.1%, about 0.15%, about 0.2%, about 0.25%, about 0.3%, about 0.35%, about 0.4%, about 0.45%, or about 0.5% of the total weight.

[0040] In some embodiments, the content of the rifamycin-quinolidinone coupled molecule is 0.5% by weight.

[0041] Polyethylene glycol 400 and polyethylene glycol 3350 The solubility of the rifamycin-quinolidinone coupling molecule is relatively low, and in order to obtain an ointment with suitable drug loading capacity and stability, the present application provides an ointment base comprising a combination of polyethylene glycol 400 and polyethylene glycol 3350.

[0042] In some embodiments, the content of polyethylene glycol 400 is 55% to 65% by weight. For example, polyethylene glycol 400 can be present in an amount of about 55%, about 56%, about 57%, about 58%, about 59%, about 60%, about 61%, about 62%, about 63%, about 64%, or about 65% of the total weight.

[0043] In some embodiments, the content of polyethylene glycol 400 is 55% to 60% by weight. For example, polyethylene glycol 400 can be present in an amount of about 55%, about 56%, about 57%, about 58%, about 59%, or about 60% of the total weight.

[0044] In some embodiments, the content of polyethylene glycol 400 is 56% to 58% by weight. For example, polyethylene glycol 400 can be present in an amount of about 56%, about 56.1%, about 56.2%, about 56.3, about 56.4, about 56.5, about 56.6%, about 56.7%, about 56.8%, about 56.9%, about 57%, about 57.1%, about 57.2%, about 57.3, about 57.4, about 57.5, about 57.6%, about 57.7%, about 57.8%, about 57.9%, or about 58% of the total weight.

[0045] In some embodiments, the content of polyethylene glycol 3350 is 25% to 35% by weight. For example, the content of polyethylene glycol 3350 can be about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, or about 35% of the total weight.

[0046] In some embodiments, the content of polyethylene glycol 3350 is 28% to 33% by weight. For example, the content of polyethylene glycol 3350 can be about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, or about 35% of the total weight.

[0047] In some embodiments, the content of polyethylene glycol 3350 is 30% to 32% by weight. For example, polyethylene glycol 3350 can be present in an amount of about 30%, about 30.1%, about 30.2%, about 30.3%, about 30.4, about 30.5, about 30.6%, about 30.7%, about 30.8%, about 30.9%, about 31%, about 31.1%, about 31.2%, about 31.3, about 31.4, about 31.5, about 31.6%, about 31.7%, about 31.8%, about 31.9%, or about 32% of the total weight.

[0048] Propylene glycol The ointment of the present application not only considers the stability of the formulation, but also improves the relatively hard spreading caused by the polyethylene glycol base by adding a certain percentage of propylene glycol, thereby improving the patient or user's experience. In some embodiments, the propylene glycol content is 8% to 12% by weight. For example, propylene glycol may be present in an amount of about 8%, about 8.5%, about 9%, about 9.5%, about 10%, about 10.5%, about 11%, about 11.5%, or about 12% of the total weight. Specifically, propylene glycol may be present in an amount of about 10% of the total weight.

[0049] solubilizer To address the low solubility of the rifamycin-quinolidinone coupling molecule and its potentially adverse tendency toward API precipitation or aggregation, a suitable solubilizer is also added to the ointment components of the present application. In some embodiments, the solubilizer is present in an amount of 0.1% to 1% by weight. For example, the solubilizer may be present in an amount of about 0.1%, about 0.15%, about 0.2%, about 0.25%, about 0.3%, about 0.35%, about 0.4%, about 0.45%, about 0.5%, about 0.55%, about 0.6%, about 0.65%, about 0.7%, about 0.75%, about 0.8%, about 0.85%, about 0.9%, about 0.95%, or about 1% of the total weight.

[0050] In some embodiments, the solubilizer is present in an amount of 0.3% to 0.7% by weight, e.g., about 0.3%, about 0.31%, about 0.32%, about 0.33%, about 0.34%, about 0.35%, about 0.36%, about 0.37%, about 0.38%, about 0.39%, about 0.4%, about 0.41%, about 0.42%, about 0.43%, about 0.44%, about 0.45%, about 0.46%, about 0.47%, about 0.48%, or about 0.49% of the total weight. , about 0.5%, about 0.51%, about 0.52%, about 0.53%, about 0.54%, about 0.55%, about 0.56%, about 0.57%, about 0.58%, about 0.59%, about 0.6%, about 0.61%, about 0.62%, about 0.63%, about 0.64%, about 0.65%, about 0.66%, about 0.67%, about 0.68%, about 0.69%, or about 0.7%.

[0051] In some embodiments, the solubilizing agent content can include triethanolamine. For example, the rifamycin-quinolizinon coupled molecule ointments described herein may contain about 0.3%, about 0.31%, about 0.32%, about 0.33%, about 0.34%, about 0.35%, about 0.36%, about 0.37%, about 0.38%, about 0.39%, about 0.4%, about 0.41%, about 0.42%, about 0.43%, about 0.44%, about 0.45%, about 0.46%, about 0.47%, about 0.48% or about 0.50% of the total mass. %, about 0.49%, about 0.5%, about 0.51%, about 0.52%, about 0.53%, about 0.54%, about 0.55%, about 0.56%, about 0.57%, about 0.58%, about 0.59%, about 0.6%, about 0.61%, about 0.62%, about 0.63%, about 0.64%, about 0.65%, about 0.66%, about 0.67%, about 0.68%, about 0.69%, or about 0.7% triethanolamine.

[0052] antioxidants The storage stability and oxidative degradation of the rifamycin-quinolidinone coupled molecule ointments described herein can be improved by adding an antioxidant. In some embodiments, the antioxidant is present in an amount of 0.3% to 1% by weight. For example, the antioxidant may be present in an amount of about 0.3%, about 0.35%, about 0.4%, about 0.45%, about 0.5%, about 0.55%, about 0.6%, about 0.65%, about 0.7%, about 0.75%, about 0.8%, about 0.85%, about 0.9%, about 0.95%, or about 1% of the total weight.

[0053] In some embodiments, the antioxidant is present in an amount of 0.5% to 1% by weight, for example, about 0.5%, about 0.51%, about 0.52%, about 0.53%, about 0.54%, about 0.55%, about 0.56%, about 0.57%, about 0.58%, about 0.59%, about 0.6%, about 0.61%, about 0.62%, about 0.63%, about 0.64%, about 0.65%, about 0.66%, about 0.67%, about 0.68%, about 0.69%, about 0.7%, about 0.71%, about 0.72%, about 0.73%, about 0.74%, or about 0.75% of the total weight. It may be present in an amount of about 0.75%, about 0.76%, about 0.77%, about 0.78%, about 0.79%, about 0.8%, about 0.81%, about 0.82%, about 0.83%, about 0.84%, about 0.85%, about 0.86%, about 0.87%, about 0.88%, about 0.89%, about 0.9%, about 0.91%, about 0.92%, about 0.93%, about 0.94%, about 0.95%, about 0.96%, about 0.97%, about 0.98%, about 0.99%, or about 1%.

[0054] In some embodiments, the antioxidant comprises one or more of vitamin C and vitamin E. For example, the antioxidant can be vitamin C, vitamin E, or a combination of vitamin C and vitamin E. For example, the rifamycin-quinolidinone coupled molecule ointments described herein may contain vitamin C, vitamin E, or a combination of vitamin C and vitamin E at about 0.5%, about 0.51%, about 0.52%, about 0.53%, about 0.54%, about 0.55%, about 0.56%, about 0.57%, about 0.58%, about 0.59%, about 0.6%, about 0.61%, about 0.62%, about 0.63%, about 0.64%, about 0.65%, about 0.66%, about 0.67%, about 0.68%, about 0.69%, about 100%, about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 1109%, about 1111%, about 112%, about 113%, about 114%, about 115%, about 116%, about 117%, about 118%, about 119%, about 120%, about 121%, about 122%, about 123%, about 124%, about 125%, about 126%, about 127%, about 128%, about 129%, about 130%, about 131%, about 132%, about 133%, about 134%, about 135%, about 136%, about 137%, about 138%, about 139%, about 140%, about 141%, about 142%, about 143%, about 144%, about 145%, about 146%, about 147%, about 1 %, about 0.71%, about 0.72%, about 0.73%, about 0.74%, about 0.75%, about 0.76%, about 0.77%, about 0.78%, about 0.79%, about 0.8%, about 0.81%, about 0.82%, about 0.83%, about 0.84%, about 0.85%, about 0.86%, about 0.87%, about 0.88%, about 0.89%, about 0.9%, about 0.91%, about 0.92%, about 0.93%, about 0.94%, about 0.95%, about 0.96%, about 0.97%, about 0.98%, about 0.99%, or about 1%.

[0055] In some embodiments, the content of vitamin C and / or vitamin E is 0.3% to 0.5% by weight. For example, vitamin C can be present in an amount of about 0.3%, about 0.31%, about 0.32%, about 0.33%, about 0.34%, about 0.35%, about 0.36%, about 0.37%, about 0.38%, about 0.39%, about 0.4%, about 0.41%, about 0.42%, about 0.43%, about 0.44%, about 0.45%, about 0.46%, about 0.47%, about 0.48%, about 0.49%, or about 0.5% of the total weight. and / or Vitamin E may be present in an amount of about 0.3%, about 0.31%, about 0.32%, about 0.33%, about 0.34%, about 0.35%, about 0.36%, about 0.37%, about 0.38%, about 0.39%, about 0.4%, about 0.41%, about 0.42%, about 0.43%, about 0.44%, about 0.45%, about 0.46%, about 0.47%, about 0.48%, about 0.49%, or about 0.5% of the total mass.

[0056] Method of preparing the ointment The method for preparing a rifamycin-quinolidinone coupled molecule ointment described herein includes heat-melting polyethylene glycol 3350, propylene glycol, a solubilizer, an antioxidant, and polyethylene glycol 400 to obtain a first mixture, wherein a portion of the regulated amount of polyethylene glycol 400 may be added during heat-melting, or the entire regulated amount of polyethylene glycol 400 may be added. In some cases, the heat-melting is performed at a temperature of 70°C or higher. For example, the heat-melting can be performed at a temperature of about 70°C, about 71°C, about 72°C, about 73°C, or higher. The preparation method described herein further includes mixing the rifamycin-quinolidinone coupled molecule with a first mixture containing all of the polyethylene glycol 400, and optionally the mixing step is performed at a temperature of 50°C or higher, for example, the mixing step is performed at a temperature of about 50°C, about 51°C, about 52°C, about 53°C, or higher; or the preparation method described herein further includes mixing and dissolving the rifamycin-quinolidinone coupled molecule with the remainder of the polyethylene glycol 400 from the heat-melting step to obtain a first mixture, and then mixing the obtained solution with the first mixture, and the mixing and dissolving, as well as the mixing with the first mixture, are all performed at a temperature of 50°C or higher, for example, the mixing and dissolving, and the mixing with the first mixture can be performed at a temperature of about 50°C, about 51°C, about 52°C, about 53°C, or higher.

[0057] Without being bound by theory, the following examples are intended only to illustrate the ointment of the present application, its method of preparation, its use, etc., and are not intended to limit the scope of the present application.

[0058] example Example 1 Selection of ointment base Ointments are a preferred choice as a relatively safe topical formulation, because they are easy to use and have a targeted effect on skin and skin tissue infections. Commonly used ointment bases include PEG400 (i.e., polyethylene glycol 400). Considering the relatively low solubility of rifamycin-quinolidinone coupling molecules, the base must be selected to ensure relatively good solubility of the drug in the base, thereby increasing the drug loading capacity of the formulation.

[0059] Experimental procedure: Approximately 340 mg of API was gradually and slowly added to approximately 5 g of PEG400 (in a 40°C water bath), and the mixture was dissolved by magnetic stirring. The orange-red mixture gradually became viscous and could no longer be stirred. Approximately 7 g of PEG400 was gradually added and stirred for an additional 20 minutes. The resulting mixture was divided into two portions, one of which was kept in a 40°C water bath and the other was cooled to room temperature. Both portions were filtered through a PES water membrane (13 mm, 0.45 μm), and the filtrate was collected for content analysis.

[0060] Approximately 400 mg of API was slowly and gradually added to approximately 5 g of propylene glycol (in a 40°C water bath), and the mixture was stirred to dissolve. The orange-red mixture gradually became viscous and could no longer be stirred. Approximately 7 g of PEG 400 was slowly added and stirred for an additional 20 minutes. The resulting mixture was divided into two portions, one of which was kept in a 40°C water bath and the other was cooled to room temperature. Both portions were filtered through a PES water membrane (13 mm, 0.45 μm), and the filtrate was collected for content analysis.

[0061] Four filtrates were tested for drug content as shown in Table 1 below. [Table 1]

[0062] As shown in the table above, the solubility of the rifamycin-quinolidinone coupled molecule in PEG400 at 40°C exceeds the formulation specification (specification: 0.5%, i.e., 5 mg / g), which is better than the solubility at room temperature and significantly better than other bases such as propylene glycol. Therefore, PEG400 was selected as the primary base.

[0063] However, when PEG400 was used as the sole base for the ointment, the ointment was relatively hard and had poor spreading properties. After several experiments, different proportions of propylene glycol were added to the base to obtain the following formulations for comparative testing. The formulation compositions are shown in Table 2 below: [Table 2]

[0064] Experimental Procedure: Excipients were weighed according to the formulation and processed using a one-pot method. All excipients from each batch were added simultaneously and heated in a water bath at 65-73°C with magnetic stirring. The mixture was slowly stirred and then melted. After melting, the mixture was stirred at 1500 rpm for 10 minutes, then stirred at 800 rpm at room temperature and allowed to cool and solidify.

[0065] The above base formulations with different ratios were left in a refrigerator at 2-8°C overnight. The next day, the formulations were removed and returned to room temperature. The appearance and microscopic images of the products were observed to evaluate their spreadability. By comparison, Formulations 1, 2, and 3 exhibited a waxy appearance, with Formulation 3 in particular exhibiting waxy solid-liquid separation; Formulations 4, 5, 6, and 7 exhibited a snowflake-like structure and were observed to gradually thin. The spreadability of the corresponding seven formulas transitioned from spreadable to easily and uniformly spreadable. Microscopic observation revealed that the bases of Formulations 4 and 5 exhibited a relatively more uniform distribution, indicating that they were more suitable as bases.

[0066] Tests have shown that the advantages of using propylene glycol include: 1. Propylene glycol has a lower viscosity than PEG400, is miscible with PEG400, and can be melt-mixed with PEG3350, significantly improving the sensory properties of relatively hard polyethylene glycol hydrophilic ointments; 2. Propylene glycol, a commonly used moisturizer in topical formulations, can maintain the consistency of the ointment; 3. Propylene glycol has specific antibacterial activity. In the examples of this application, a comparison is made with ointment bases, and the products of the examples of this application can solve the problems that existing ointments have a relatively hard, waxy texture and poor spreadability.

[0067] Example 2 Selection of oxidizing agent and its amount This example provides a rifamycin-quinolidinone coupled molecule ointment, the components of which, based on mass percentage, include the rifamycin-quinolidinone coupled molecule in an amount of 0.5% of the total mass, polyethylene glycol 400 in an amount of 56% to 58% of the total mass, polyethylene glycol 3350 in an amount of 30% to 32% of the total mass, propylene glycol in an amount of 10% of the total mass, vitamin C in an amount of 0 to 0.5% of the total mass, vitamin E in an amount of 0 to 0.5% of the total mass, and BHT in an amount of 0 to 0.5% of the total mass.

[0068] The rifamycin-quinolidinone coupled molecule ointment of this example was prepared by the following method: Polyethylene glycol 3350, propylene glycol, antioxidant, and polyethylene glycol 400 were added to a container, and the mixture was heated and melted in a water bath at 70-73°C and slowly mechanically stirred at 1000 rpm until a clear solution was obtained. The mixture was stirred for an additional 10 minutes and mixed to obtain a first mixture. The rifamycin-quinolidinone coupled molecule was mixed with the first mixture in a water bath at 50-53°C, and the mixture was mechanically stirred at 1000 rpm for approximately 15 minutes until no rifamycin-quinolidinone coupled molecule powder was observed with the naked eye. The mixture was continuously stirred and cooled to 40-43°C to obtain a rifamycin-quinolidinone coupled molecule ointment.

[0069] All ointments of rifamycin-quinolidinone coupling molecules, formulations 8 to 12, were prepared using this method, except for the slight differences in the formulation composition of each excipient.

[0070] During the preparation process, the rifamycin-quinolidinone coupling molecule undergoes oxidative decomposition upon exposure to air, resulting in the generation of oxidized impurities. The structural changes resulting from the reaction are shown in Figure 1. To reduce the oxidative decomposition rate of the rifamycin-quinolidinone coupling molecule, several antioxidants, such as butylated hydroxytoluene (BHT), vitamin C, and vitamin E, were investigated, both alone and in combination, along with their dosages. The formulations are shown in Table 3. [Table 3]

[0071] According to the formulation in the above table, rifamycin-quinolidinone coupling molecule ointment was prepared and related substances were examined. The results are shown in Table 4 below: [Table 4]

[0072] As shown in the table above, after Formula 8 was left standing at room temperature for 7 days, the oxidized impurities increased from 0.37% to 3.55%, and the total impurities also increased significantly, indicating that BHT was unable to reduce the generation of oxidized impurities due to decomposition. However, the addition of antioxidant VC (VC stands for vitamin C) significantly improved the stability of the formula and inhibited oxidative degradation. Among these formulas, Formula 11 showed the smallest increase in both oxidized impurities and total impurities, indicating that the combination of antioxidants, 0.5% VC + 0.5% VE (VE stands for vitamin E), had a better effect on improving the stability of the formula. In the examples of this application, the type and proportion of antioxidants were compared, and the product of the example of this application can improve the problem of existing ointments undergoing decomposition to generate oxidized impurities and having poor formulation stability.

[0073] Example 3 Screening and examination of solubilizers The ointment prepared in Example 2 was subjected to microscopic examination for comparison, and the results are shown in Figure 2. The material of Formulation 11 had a tendency for red API precipitation or aggregation after 5 months of storage at 2-8°C. Although the content uniformity was acceptable, the RSD was relatively high (5.5%), which may be attributed to API precipitation, resulting in an insufficient content uniformity value.

[0074] Due to the insufficient solubility of the API itself, triethanolamine was added as a solubilizer in the examples of this disclosure. One carboxylic acid group and three phenolic hydroxyl groups in the structure of the rifamycin-quinolidinone coupling molecule are H +Triethanolamine provides a lone electron pair, forming an ion-pair complex with the organic acid group of the rifamycin-quinolidinone coupling molecule. The ion-pair complex has a low lattice energy, making the rifamycin-quinolidinone coupling molecule more easily soluble in solvents. Meanwhile, triethanolamine exhibits basicity and is used to adjust the pH of the ointment to neutral. The acid-base properties of the ointment were investigated. The amount of triethanolamine added was calculated in advance from the amounts of API and vitamin C added. Theoretically, triethanolamine provides a lone electron pair, is a Lewis base, and can form salts with carboxylic acids or Lewis acids at room temperature without catalyst. The reaction equation is N-(CH2CH2OH)3 + HCOOH = HCOO - +[HN-(CH2CH2OH)3] + One mole of vitamin C in the ointment is equivalent to one mole of H + When one carboxylic acid group and three phenolic hydroxyl groups in the API structure are H + Taking 100g of ointment as an example, 0.4235g of triethanolamine is required for 0.5g of vitamin C in the neutralization formula, and the H provided by one carboxylic acid group and three phenolic hydroxyl groups in the API. + 0.26 g of triethanolamine was required to neutralize the ethanolamine. Therefore, the amount of triethanolamine added should theoretically be less than 0.4235 g + 0.26 g = 0.6835 g. Based on this calculation, gradients of 0.3%, 0.5%, and 0.7% of the triethanolamine formulation addition were established.

[0075] Based on formulation 11, 0.3%, 0.5%, and 0.7% triethanolamine were added to obtain formulations 13–15, respectively, to study the effects of different formulations on the pH, related substances, and properties of the ointment. [Table 5]

[0076] As shown in the table above, the addition of triethanolamine was able to adjust the pH of the ointment from slightly acidic to neutral. [Table 6]

[0077] The structure of the acid degradation impurity was shown as the following formula: [ka] The batch stability results show that when triethanolamine was added at 0.3%, 0.5%, and 0.7%, the total impurities in the samples after 3 months of storage at 2-8°C were 1.32%, 1.12%, and 1.02%, respectively, indicating improved chemical stability compared to Formulation 11, which contained only antioxidants and had a total impurity level of 1.69% (Table 7). Furthermore, as can be seen from Table 6, no precipitation or aggregation of the API was observed in the three batches stored at 5°C for 3 months, indicating improved physical stability and demonstrating the solubilization effect.

[0078] Example 4 Preparation of ointment of rifamycin-quinolidinone coupling molecule This example provides a rifamycin-quinolidinone coupled molecule ointment, the components of which, by mass percentage, include the rifamycin-quinolidinone coupled molecule in an amount of 0.5% of the total mass, polyethylene glycol 400 in an amount of 56% of the total mass, polyethylene glycol 3350 in an amount of 32% of the total mass, propylene glycol in an amount of 10% of the total mass, triethanolamine in an amount of 0.5% of the total mass, vitamin C in an amount of 0.5% of the total mass, and vitamin E in an amount of 0.5% of the total mass.

[0079] The rifamycin-quinolidinone coupled molecule ointment of this example was prepared by the following method: Add polyethylene glycol 3350 (abbreviated as PEG3350), propylene glycol, solubilizer, antioxidant, and polyethylene glycol 400 (abbreviated as PEG400) to a container, heat the mixture in a water bath at 70-73°C to melt it, and slowly stir the mixture at a rotation speed of 1000 rpm until a clear solution is obtained; then stir and mix the mixture for another 10 minutes to obtain a mixture; The rifamycin-quinolidinone coupled molecule was mixed with the mixture in a water bath at 50-53°C, and the mixture was mechanically stirred at 1000 rpm for approximately 15 minutes until no rifamycin-quinolidinone coupled molecule powder was observed with the naked eye; the mixture was continuously stirred and cooled to 40-43°C to obtain a rifamycin-quinolidinone coupled molecule ointment.

[0080] All rifamycin-quinolidinone coupled molecule ointments were prepared using this method, except for slight variations in the formulation of each excipient.

[0081] Example 5 Antibacterial activity test of rifamycin-quinolidinone coupling molecule ointment on skin surface Staphylococcus aureus (S. aureus) is one of the most common bacterial pathogens in humans and can cause skin and soft tissue infections (SSTIs), bacteremia, osteomyelitis, and other disseminated infections. The penicillin-binding protein PBP2a is a major cause of drug resistance in S. aureus. PBP2a has low binding capacity for β-lactam antibiotics and can maintain the stability of bacterial peptidoglycan in the presence of such antibiotics, thereby conferring drug resistance to the bacterium. Strains containing PBP2a are called methicillin-resistant Staphylococcus aureus (MRSA). S. aureus NRS 384 is one such strain and has been noted to cause infections in healthcare settings in the United States, primarily associated with skin and soft tissue infections. The rifamycin-quinolidinone ointment, prepared by TenNor Therapeutics (Suzhou) Limited, is an ointment formulation intended for the treatment of S. aureus infections. In this experiment, a mouse skin infection model was established using S. aureus NRS 384 to evaluate the antibacterial activity of the rifamycin-quinolidinone ointment on the skin surface.

[0082] Experimental Design: Six groups were set up in this experiment, including a blank ointment group, a bactroban ointment group, and three dose groups of rifamycin-quinolidinone coupling molecule ointment (low, medium, and high dose groups). Each group contained eight animals, and the specific grouping is shown in Table 7. [Table 7]

[0083] Test Results: 1) Counting results of inoculated bacterial solution: The CFU count results of the inoculated bacterial solution were as follows: the actual inoculum concentration was 3.85E+09 CFU / mL, and the inoculum dose for each mouse was 3.85E+07 CFU. [Table 8]

[0084] 2) Counting results of bacterial load on mouse skin: [Table 9]

[0085] The results of this pharmacodynamic experiment showed that the bacterial load on the skin of the blank ointment group was 9.154 ± 0.273 lg. Using S. aureus NRS 384, we were able to establish a stable skin infection model (Table 9, Figure 3). Compared to the blank ointment group, the bacterial load on mouse skin was reduced by 2.119 logs after treatment with 30 mg / mouse of Bactroban ointment (2% mupirocin), demonstrating the bactericidal effect of Bactroban ointment. Animals treated with 0.5%, 0.05%, and 0.005% rifamycin-quinolidinone coupled molecule ointments showed a reduction in bacterial burden on the skin of 4.718 Log, 3.100 Log, and 1.063 Log, respectively, indicating that the 0.5% and 0.05% high-dose rifamycin-quinolidinone coupled molecule ointments showed good bactericidal effects in this skin infection model, with highly significant differences from the animals in the blank ointment group.

[0086] Testing conclusion: In this study, we established a stable mouse skin infection model using S. aureus NRS 384 to evaluate the antibacterial activity of rifamycin-quinolidinone ointment on the skin surface. The results show that rifamycin-quinolidinone ointment can effectively reduce bacterial load on mouse skin and exhibits dose-dependent bactericidal activity.

Claims

1. An ointment of a rifamycin-quinolidinone coupled molecule, comprising the following components: comprising a rifamycin-quinolidinone coupled molecule of Formula I, polyethylene glycol 400, polyethylene glycol 3350, propylene glycol, a solubilizer, and an antioxidant; 【Chemical 1】 Based on a total mass percentage of 100% the content of the rifamycin-quinolidinone coupled molecule is 0.05% by mass to 0.7% by mass; The content of polyethylene glycol 400 is 55% by mass to 65% by mass, The content of polyethylene glycol 3350 is 25% by mass to 35% by mass, The content of propylene glycol is 8% by mass to 12% by mass, The content of the solubilizer is 0.1% by mass to 1% by mass, and The content of the antioxidant is 0.3% by mass to 1% by mass. Ointment of rifamycin-quinolidinone coupling molecule.

2. The ointment of claim 1, wherein the content of the rifamycin-quinolidinone coupled molecule is 0.3% by mass to 0.7% by mass.

3. The ointment of claim 1, wherein the content of the rifamycin-quinolidinone coupled molecule is 0.05% by mass to 0.5% by mass.

4. The ointment of any one of claims 1 to 3, wherein the content of the rifamycin-quinolidinone coupled molecule is 0.5% by mass.

5. The rifamycin-quinolidinone coupled molecule ointment according to any one of claims 1 to 4, wherein the content of polyethylene glycol 400 is 55% by mass to 60% by mass.

6. The ointment of any one of claims 1 to 5, wherein the content of polyethylene glycol 400 is 56% by mass to 58% by mass.

7. The rifamycin-quinolidinone coupled molecule ointment according to any one of claims 1 to 6, wherein the content of polyethylene glycol 3350 is 28% by mass to 33% by mass.

8. The ointment of any one of claims 1 to 7, wherein the content of polyethylene glycol 3350 is 30% by mass to 32% by mass.

9. The rifamycin-quinolidinone coupled molecule ointment according to any one of claims 1 to 8, wherein the content of the solubilizer is 0.3% by mass to 0.7% by mass.

10. The rifamycin-quinolidinone coupled molecule ointment of any one of claims 1 to 9, wherein the solubilizing agent comprises triethanolamine.

11. The rifamycin-quinolidinone coupled molecule ointment according to any one of claims 1 to 10, wherein the content of the antioxidant is 0.5% by mass to 1% by mass.

12. The rifamycin-quinolidinone coupled molecule ointment according to any one of claims 1 to 11, wherein the antioxidant comprises one or more of vitamin C and vitamin E.

13. The ointment of claim 12, wherein the content of vitamin C and / or vitamin E is 0.3% by mass to 0.5% by mass.

14. The components of the rifamycin-quinolidinone coupling molecule ointment are, based on a total mass percentage of 100%, 0.5% by weight of the rifamycin-quinolidinone coupled molecule; 56% to 58% by weight of polyethylene glycol 400, 30% to 32% by weight of polyethylene glycol 3350, 10% by weight of propylene glycol, 0.3% to 0.7% by weight of triethanolamine, 0.3% to 0.5% by weight of vitamin C, and 0.3% to 0.5% by weight of vitamin E An ointment of the rifamycin-quinolidinone coupled molecule according to any one of claims 1 to 13, comprising:

15. heat-melting and mixing polyethylene glycol 3350, propylene glycol, the solubilizer, the antioxidant, and a portion of polyethylene glycol 400 to obtain a first mixture; Mixing and dissolving the rifamycin-quinolidinone coupling molecule and the remainder of polyethylene glycol 400 to obtain a first drug solution; and mixing the first drug solution with the first mixture to obtain an ointment of the rifamycin-quinolidinone coupled molecule; A method for preparing an ointment of the rifamycin-quinolidinone coupled molecule according to any one of claims 1 to 14, comprising:

16. 16. The method according to claim 15, wherein in the step of mixing and dissolving the rifamycin-quinolidinone coupled molecule and the remainder of polyethylene glycol 400 to obtain the first drug solution, the mixing and dissolving is carried out at a temperature of 50°C.

17. The preparation method according to claim 15, wherein in the step of mixing the first chemical liquid and the first mixture, the mixing is carried out at a temperature of 50 to 53°C.

18. Heating, melting, and mixing polyethylene glycol 3350, propylene glycol, the solubilizer, the antioxidant, and polyethylene glycol 400 to obtain a first mixture; and mixing the rifamycin-quinolidinone coupled molecule with the first mixture to obtain an ointment of the rifamycin-quinolidinone coupled molecule; A method for preparing an ointment of the rifamycin-quinolidinone coupled molecule according to any one of claims 1 to 14, comprising:

19. The preparation method according to claim 15 or 18, wherein the heat melting is carried out at a temperature of 70 to 73°C.

20. 19. The method of claim 18, wherein in the step of mixing the rifamycin-quinolidinone coupled molecule with the first mixture, the mixing is performed at a temperature of 50°C.

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