LIQUID FORMULATION OF XYLOPIA AETHIOPICA EXTRACT FOR PAIN MANAGEMENT IN DIABETIC NEUROPATHY AND RELATED METHODS

Abstract

ABSTRACT A liquid formulation of an acetone extract of Xylopia aethiopica (XA-LF) is developed for treating diabetic neuropathy. The formulation includes an acetone extract of dried fruits of Xylopia aethiopica and a preservative. The preparation involves extracting the dried fruits using acetone, concentrating the extract, sterilizing it, and adding a preservative. The method for treating diabetic neuropathy involves administering the XA-LF to a subject, which may improve pain threshold and reduce hyperalgesia and allodynia. Reference Fig 1

Specification

Description:DETAILED DESCRIPTION
[0015] The technical area of this process involves the development of a liquid formulation derived from the acetone extract of Xylopia aethiopica, known as XA-LF, which is intended for the treatment of diabetic neuropathy. Diabetic neuropathy is a common complication of diabetes, characterized by nerve damage that leads to symptoms such as pain, tingling, and numbness, particularly in the extremities. The formulation of XA-LF aims to address these symptoms by leveraging the natural properties of Xylopia aethiopica, a plant known for its medicinal benefits.

[0016] The active constituents, which may include flavonoids and phenolic compounds, can provide anti-inflammatory and analgesic effects, contributing to the formulation's potential as a treatment for diabetic neuropathy. The integration of the preservative into the acetone extract may be a key aspect of the formulation process, ensuring that the XA-LF is both stable and effective for therapeutic use.

[0017] The process may begin with the collection and authentication of the dried fruits of Xylopia aethiopica, which may be conducted by an organization such as NISCAIR, New Delhi.

[0018] Liquid formulation of Xylopia aethiopica extract (XA-LF), the administration of the formulation to subjects suffering from diabetic neuropathy may be considered. The XA-LF may comprise active constituents, including flavonoids and phenolic compounds, which may provide potent anti-inflammatory and analgesic effects. The formulation may be administered orally at doses of 250 mg/kg and 500 mg/kg to diabetic mice, potentially offering a promising alternative treatment for diabetic neuropathy. The active constituents of the extract may be responsible for the observed effects, which may include improved pain threshold and reduced thermal and cold hyperalgesia, as well as reduced thermal and cold allodynia in the subjects. The final XA-LF may be formulated to deliver these effects effectively to diabetic mice, potentially making it a viable option for managing pain in diabetic neuropathy patients. The potential of the XA-LF to treat diabetic neuropathy may be attributed to its active constituents, which may offer a natural, cost-effective therapy with fewer side effects compared to conventional pharmacological treatments.

[0019] FIG. 1 is a flowchart illustrating a method in step 100 for extracting the dried fruits of Xylopia aethiopica using a solvent extraction process with acetone to obtain the acetone extract, according to an embodiment. At step 100, the dried fruits of Xylopia aethiopica may be subjected to a solvent extraction process. This process may involve the use of Soxhlet extraction with 80% acetone, which can be conducted over a period of 8-10 hours. The purpose of this extraction process may be to obtain the acetone extract from the dried fruits. The Soxhlet extraction method may be chosen for its ability to efficiently extract the desired compounds from the plant material. The acetone extract obtained from this process may contain active constituents, such as flavonoids and phenolic compounds, which may provide anti-inflammatory and analgesic effects. These effects may be beneficial in the treatment of diabetic neuropathy, as they may help in reducing pain and inflammation. The extraction process may be a key step in the preparation of the liquid formulation of Xylopia aethiopica extract (XA-LF), as it may determine the concentration and efficacy of the active constituents in the final product.

[0020] At step 102, the process may involve concentrating the acetone extract. This concentration may be achieved through the use of a rotary evaporator, which can operate at a temperature below 50°C. The rotary evaporator may facilitate the removal of the solvent, thereby concentrating the extract. This step may be significant in ensuring that the active constituents of the extract are preserved and concentrated, which may enhance the efficacy of the final formulation. The use of a rotary evaporator may provide a controlled environment for the concentration process, potentially minimizing the degradation of sensitive compounds within the extract. This step may be integral to the preparation of the liquid formulation, as it may ensure that the active constituents are present in sufficient quantities to exert their therapeutic effects. The concentrated extract may then be subjected to further processing steps, such as sterilization and the addition of preservatives, to obtain the final liquid formulation.

[0021] In step 104, the process of sterilizing the concentrated acetone extract may be undertaken. This step may involve filtering the concentrated acetone extract through a 0.22 µm filter, which can serve as a sterilization method. The use of a 0.22 µm filter may be intended to remove any microbial contaminants, thereby ensuring the sterility of the extract. This action may be significant in maintaining the purity and safety of the final liquid formulation of the acetone extract of Xylopia aethiopica (XA-LF). The sterilization process may be aligned with the goal of preparing a formulation that is suitable for therapeutic use, particularly in treating diabetic neuropathy. The sterilized extract may then be ready for subsequent steps, such as the addition of preservatives, to complete the formulation process. This step may be integral to the overall method of preparing the XA-LF, as it may ensure that the extract is free from contaminants that could compromise its efficacy or safety when administered to subjects.

[0022] In the context of Step 106, the process may involve the addition of a preservative to the sterilized acetone extract to obtain the XA-LF. This step may be significant in ensuring the stability and longevity of the liquid formulation. The preservative, identified as sodium benzoate, may be added at a concentration of 0.015%, which may serve to inhibit microbial growth and maintain the integrity of the formulation. The acetone extract, having undergone prior processes of extraction, concentration, and sterilization, may be combined with sodium benzoate to form the final XA-LF. This formulation may then be prepared for subsequent administration. The addition of sodium benzoate may be a key step in the formulation process, as it may ensure that the XA-LF remains effective and safe for use in treating diabetic neuropathy. The potential actions described in this step may align with the overall goal of creating a stable and efficacious liquid formulation for therapeutic use.

[0023] In step 108, the process may involve administering the XA-LF to subjects experiencing diabetic neuropathy. The administration can be performed orally, with potential doses of 250 mg/kg or 500 mg/kg. The XA-LF may contain active constituents, such as flavonoids and phenolic compounds, which can provide anti-inflammatory and analgesic effects. These effects may contribute to the treatment of diabetic neuropathy by potentially improving the pain threshold and reducing symptoms such as thermal hyperalgesia, cold hyperalgesia, thermal allodynia, and cold allodynia in the subjects. The administration of the XA-LF may be aimed at delivering these active constituents effectively to the diabetic mice, thereby offering a promising alternative treatment for managing pain associated with diabetic neuropathy.

[0024] The formulation's potential efficacy in reducing hyperalgesia and allodynia may be attributed to the active constituents' properties, which can provide a natural and cost-effective therapy with fewer side effects compared to conventional pharmacological treatments.

Examples
[0025] Example 1: Phytochemical Screening:
The phytochemical analysis of the Xylopia aethiopica extract was conducted to identify the presence of alkaloids, flavonoids, phenolic compounds, tannins, and saponins, all known for their therapeutic properties.

[0026] Induction of Diabetic Neuropathy in Mice- Animal Model:
Materials and Methods
The study utilized Swiss albino mice (25-30 g), maintained under standard laboratory conditions with free access to food and water. All procedures were approved by the Institutional Animal Ethical Committee (IAEC). Diabetic neuropathy was induced by administering streptozotocin (STZ) intraperitoneally at a dose of 40 mg/kg on days 1 and 3, in conjunction with a high-fat diet (HFD) for 14 days. Mice with fasting blood glucose levels above 150 mg/dL on day 14 were considered diabetic. The liquid formulation (XA-LF) was administered orally at doses of 250 mg/kg and 500 mg/kg from day 20 to day 42. Pain perception was assessed at various time points using methods such as tail flick latency, thermal and cold allodynia, and thermal hyperalgesia.

• Diabetes Induction:
DN was induced by administering streptozotocin (STZ) at a dose of 40 mg/kg intraperitoneally on days 1 and 3, in combination with a high-fat diet (HFD) for 14 days. Mice with fasting blood glucose levels over 150 mg/dl by day 14 were considered diabetic.
• XA-LF Treatment:
The liquid formulation of Xylopia aethiopica was administered orally from day 20 to day 42. Pain perception was assessed using methods such as tail flick latency, thermal allodynia, cold allodynia, and thermal hyperalgesia on various days (days 14, 21, 28, 35, and 42).

Animal Groups
• Group I: Control group (non-diabetic), administered citrate buffer and a normal diet.
• Group II: Diabetic control group, administered STZ and HFD.
• Group III: XA-LF treated group (250 mg/kg), administered STZ + HFD followed by XA-LF (250 mg/kg).
• Group IV: XA-LF treated group (500 mg/kg), administered STZ + HFD followed by XA-LF (500 mg/kg).

[0027] The formulation was administered orally for a period of 21 days, and pain perception was assessed using the following methods at baseline (Day 14), and on Days 21, 28, 35, and 42. Results are summarized in table 1 below:

[0028] Table 1: Results
Parameter Control Group (Citrate Buffer) Diabetic Control (STZ + HFD) XA-LF (250 mg/kg) XA-LF (500 mg/kg)
Baseline Tail Flick Latency (s) 5.0 ± 0.2 2.5 ± 0.3* 3.0 ± 0.4* 4.0 ± 0.3**
Tail Flick Latency (Day 21) 5.0 ± 0.2 2.5 ± 0.3* 3.5 ± 0.3C** 4.5 ± 0.2**
Cold Allodynia Response (s) 5.0 ± 0.2 1.5 ± 0.2* 3.0 ± 0.3** 4.0 ± 0.4**
Thermal Allodynia Response (s) 5.0 ± 0.2 2.0 ± 0.3* 3.0 ± 0.3** 4.5 ± 0.3**

Values are expressed as mean ± standard deviation (SD).
* indicates significant difference compared to control group (p < 0.05).
** indicates significant difference compared to diabetic control group (p < 0.05).

[0029] Phytochemical analysis of the acetone extract confirmed the presence of flavonoids, phenolic acids, tannins, and saponins, all of which contribute to the observed therapeutic effects. The percentage yield of the acetone extract was calculated to be 9.25%, resulting in a dark brown viscous liquid. This formulation demonstrates significant efficacy in alleviating the pain and sensory abnormalities associated with diabetic neuropathy, offering a promising, natural, and cost-effective alternative to conventional pharmacological treatments.

[0030] STABILITY DATA:
Stability studies were conducted to assess the formulation's ability to retain its physical characteristics (such as appearance, viscosity, and pH), the concentration of active ingredients (flavonoids and phenolic compounds), and its microbiological stability over time. These tests were performed under various environmental conditions, including room temperature (25°C ± 2°C, 60% ± 5% RH), refrigerated conditions (5°C ± 3°C), and elevated temperature (40°C ± 2°C, 75% ± 5% RH).

[0031] The formulation was evaluated at specific time intervals-0, 3, 6, and 12 months-to monitor any changes in its properties.

[0032] Table 2: Stability Data
Parameter Time Room Temperature (25°C ± 2°C,
60% ± 5% RH) Refrigerated (5°C ± 3°C) Elevated Temperature
(40°C ± 2°C, 75% ± 5% RH)
Physical Appearance 0 Dark brown, viscous; no separation or precipitation. Dark brown, viscous; no separation. Dark brown, viscous; no separation.
3 No change. No change. Slight darkening; no precipitation.
6 No change. No change. Slight darkening; no precipitation.
12 Slight darkening; no separation or precipitation. No change. Significant darkening; no precipitation.
pH 0 5.9 5.9 5.9
3 5.8 5.9 5.7
6 5.8 5.9 5.6
12 5.7 5.8 5.5
Viscosity 0 Normal viscosity. Normal viscosity. Normal viscosity.
3 No change. No change. Slight increase.
6 No change. No change. Mild increase.
12 Slight increase in viscosity. No change. Noticeable increase in viscosity.
Assay of Active Ingredients 0 100% 100% 100%
3 99% 99% 97%
6 98% 99% 95%
12 97% 98% 92%
Microbiological Contamination 0 No contamination detected No contamination detected No contamination detected
3 No contamination detected No contamination detected No contamination detected
6 No contamination detected No contamination detected No contamination detected
12 No contamination detected No contamination detected No contamination detected







, Claims:CLAIMS
We claim:
1. A liquid formulation of an acetone extract of Xylopia aethiopica (XA-LF) for treating diabetic neuropathy, comprising:
an acetone extract of dried fruits of Xylopia aethiopica; and
a preservative;
wherein the liquid formulation is prepared by:
extracting the dried fruits of Xylopia aethiopica using a solvent extraction process with acetone to obtain the acetone extract;
concentrating the acetone extract;
sterilizing the concentrated acetone extract; and
adding the preservative to the sterilized acetone extract to obtain the XA-LF.

2. The liquid formulation of claim 1, wherein the solvent extraction process comprises Soxhlet extraction using 80% acetone for 8-10 hours.

3. The liquid formulation of claim 1, wherein concentrating the acetone extract comprises using a rotary evaporator at a temperature below 50°C.

4. The liquid formulation of claim 1, wherein sterilizing the concentrated acetone extract comprises filtering the concentrated acetone extract through a 0.22 µm filter.

5. The liquid formulation of claim 1, wherein the preservative is sodium benzoate.

6. The liquid formulation of claim 5, wherein the sodium benzoate is added at a concentration of 0.015%.

7. The liquid formulation of claim 1, wherein the XA-LF comprises active constituents including at least one of flavonoids or phenolic compounds, and wherein the active constituents provide anti-inflammatory effects or analgesic effects.
8. A method for treating diabetic neuropathy, comprising:
preparing a liquid formulation of an acetone extract of Xylopia aethiopica (XA-LF) by:
extracting dried fruits of Xylopia aethiopica using a solvent extraction process with acetone to obtain the acetone extract;
concentrating the acetone extract;
sterilizing the concentrated acetone extract; and
adding a preservative to the sterilized acetone extract to obtain the XA-LF; and
administering the XA-LF to a subject suffering from diabetic neuropathy.

9. The method of claim 8, wherein administering the XA-LF comprises oral administration at a dose of 250 mg/kg or 500 mg/kg.

10. The method of claim 8, wherein administering the XA-LF results in at least one of improved pain threshold, reduced thermal hyperalgesia, reduced cold hyperalgesia, reduced thermal allodynia, or reduced cold allodynia in the subject.

Documents