Description:1
FORM-2
THE PATENTS ACT, 1970
(39 of 1970)
&
THE PATENTS RULES, 2003
COMPLETE
SPECIFICATION
(See section 10 and rule 13)
DoE Assisted Development of Insulin Loaded Topical Organogel Formulation for the Management of Diabetic Wound
G D Goenka University, an Indian university of Sohna Gurugram Road, Sohna, Haryana, India, 122103
THE FOLLOWING SPECIFICATION PARTICULARLY DESCRIBES THE INVENTION AND THE MANNER IN WHICH IT IS TO BE PERFORMED
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FIELD OF INVENTION:
The present invention relates to pharmaceutical science field, which aims at DoE Assisted Development and In-Vitro Characterization of Insulin Loaded Topical Organogel Formulation for the Management of Diabetic Wound.
BACK GROUND:
Diabetes, also known as diabetes mellitus, is a group of common endocrine diseases characterized by sustained high blood sugar levels. Diabetes is due to either the pancreas not producing enough insulin, or the cells of the body not responding properly to the insulin produced. Diabetes, if left untreated, leads to many health complications. Untreated or poorly treated diabetes accounts for approximately 1.5 million deaths per year.
There is no widely accepted cure for most cases of diabetes. The most common treatment for type 1 diabetes is insulin replacement therapy (insulin injections). Anti-diabetic medications such as metformin and semaglutide, as well as lifestyle modifications, can be used to prevent or respond to type 2 diabetes. Gestational diabetes normally resolves shortly after delivery.
Most medications used to treat diabetes act by lowering blood sugar levels through different mechanisms. There is broad consensus that when people with diabetes maintain tight glucose control – keeping the glucose levels in their blood within normal ranges – they experience fewer complications, such as kidney problems or eye problems. There is however debate as to whether this is appropriate and cost effective for people later in life in whom the risk of hypoglycemia may be more significant.
The physiological mechanisms behind the increased severity of diabetic wounds are not explicitly targeted by any of the current therapeutic methods, and the mechanism seemed to be the most complex and problematic in many ways, as it could be argued that it would be unlikely to transfer from temporarily diabetic humans with small wounds to permanently diabetic humans with larger injuries.
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Numerous studies have been conducted depending on the kind and extent of wounds. However, there are no effective medications that are extremely stable, less costly or have no adverse effects that can help wounds heal more quickly. Traditional wound dressings such as gauze, hydrocolloids, films, and foams are single-purpose, functioning as a physical barrier or absorbing exudates, and cannot cover all of the requirements of the complete chronic wound healing process. Along with the fact that current standard treatment methods frequently result in unintended side effects, it has become necessary to discover alternative therapeutic models that are secure or have few side effects and hazards.
Gallagher et al., 2007 carried out Dipeptidyl peptidase-IV inhibitors (DPP-4) showed MMP stimulation facilitates epithelial cell emigration, which was primarily brought on by an elevation in HMGB1, one of DPP4's targets, that has previously been shown to exhibit chemotactic effects and enhance wound closure in fibroblasts and keratinocytes as well as diabetic rats. In melanonocytes, DPP4 suppression or a subsequent rise in SDF1 synthesis has also been linked to improve squamous to fibroblast transformation, which seems to be important for tissue healing. Stimulated SDF1 on the target of the injury has also been found to promote angiogenesis by restoring normal progenitor cell recruitment (Straino et al., 2008; Ranzato et al., 2010; Marfella et al., 2012).
Sharma et al., 2022 formulate pluronic F127-tailored lecithin organogel of acyclovir and reported that organogel is a good drug delivery platform for the treatment of topical disorders due to its attributes, including biocompatibility and amphiphilic nature, which promote the dissolving of diverse medications with varying solubility characteristics and increase the permeation potential of active molecules.
Wang & Xu, 2020 worked on topical Insulin modulates the inflammatory and proliferative phases of burn wound healing in diabetes-induced rats, and the result was that topical insulin modulates burn wound healing in diabetic animals by balancing inflammation and promoting angiogenesis and the formation of elastic fibers.
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Advanced wound healing therapies: Although various wound healing therapies exist, such as growth factors, skin substitutes, and stem cell-based treatments, their efficacy and optimal application in diabetic wounds are still not well understood. Further research is needed to evaluate the effectiveness of these advanced therapies in promoting diabetic wound healing. Biofilm management: Biofilms are complex communities of microorganisms that form on the wound bed and can impede the healing process. Diabetic wounds are particularly susceptible to biofilm formation. Research is needed to develop effective strategies for biofilm detection, prevention, and treatment specific to diabetic wounds. Diabetic wounds often exhibit delayed healing compared to non-diabetic wounds. The problem statement focuses on understanding the underlying mechanisms that contribute to delayed wound healing in diabetes and developing interventions to promote timely healing. Diabetic wounds are prone to infection due to impaired immune function and the presence of elevated blood sugar levels. The problem statement aims to identify strategies for effectively preventing and managing infections in diabetic wounds, including the development of novel antimicrobial therapies and improved wound care practices
OBJECTIVE OF THE INVENTION:
1. It is an object of the invention to develop and characterize of insulin loaded topical organogel formulation for the management of diabetic wound.
2. It is another object of the invention to formulate and evaluate organogel of insulin that can serve as a topical administration for promoting enhanced wound healing in diabetic patients by providing sustained and localized delivery of drug to the wound site.
SUMMARY
The combination of a topical organogel formulation with insulin for diabetic wound healing is an area of active research. Diabetes can impair wound healing due to high blood sugar levels, which can lead to reduced blood flow, nerve damage, and
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compromised immune function. Topical insulin delivery through an organogel may offer a localized approach to enhance wound healing in diabetic individuals.
Preparing the pluronic lecithin organogel, could have a great effect on wound healing because of its great properties. Pluronic F127 is a non-ionic surfactant that belongs to a group of substances called polyethylene glycols. It is widely used in pharmaceutical formulations due to its solubilizing, surfactant, and stabilizing properties. Soy lecithin is a natural substance derived from soybeans. It has emulsifying properties and improves the stability of the formulation. Isopropyl alcohol and myristic acid integrate to form the ester known as “isopropyl myristate”. It is used as a skin penetration enhancer. Due to their bi-phasic nature and flexibility, pluronic-lecithin organogels have recently grown in favor of transdermal and topical drug delivery methods. PLOs can boost the solubility of medications and the accessibility of hydrophilic pharmaceuticals. Organogels' thermo-sensitive characteristics have attracted a lot of interest in their prospective application as innovative drug-delivery technologies. Microemulsion based organogels have good potential as carriers for the topical delivery of insulin. The results of this investigation point to the insulin organogel incorporating pluronic as a drug delivery method that was secure, stable, and economical.
BRIEF DESCRIPTION OF FIGURES
These and other features, aspects, and advantages of the present invention will become better understood when the following detailed description is read with reference to the accompanying figures in which like characters represent like parts throughout the figures, wherein:
Figure 1, illustrates a view of FTIR spectrum of insulin with pluronic f 127 for the present invention.
Fig. 2: illustrates a view of FTIR spectrum of insulin with soya lecithin for the present invention.
Fig.3: illustrates a view of FTIR spectrum of mixture (insulin, polymer and excipients) for the present invention.
Fig.4: illustrates a view of Light microscopic image of formulations at 10 X
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resolutions was shown water molecules entrapped with oil, and (b). TEM images at 1.0µm resolution was shown innermost structure of formulation for the present invention.
Fig: 5: illustrates a view of pH and rheological characteristics of organogel formulations for the present invention.
Fig: 6: illustrates a view of Percentage drug content of organogel formulations for the present invention.
Fig: 7: illustrates a view of Percentage cumulative drug release data profile for the present invention.
Fig: 8: illustrates a view of Percentage cumulative drug release profile of F1-F3 formulations (a) and F4-F6 formulations (b) for the present invention.
Fig: 9: illustrates a view of Schematic representation for preparation of Insulin Loaded Topical Organogel Formulation for the present invention.
Further, skilled artisans will appreciate that elements in the figures are illustrated for simplicity and may not have been necessarily been drawn to scale. For example, the flowcharts illustrate the method in terms of the most prominent steps involved to help to improve understanding of aspects of the present invention. Furthermore, in terms of the construction of the device, one or more components of the device may have been represented in the figures by conventional symbols, and the figures may show only those specific details that are pertinent to understanding the embodiments of the present invention so as not to obscure the figures with details that will be readily apparent to those of ordinary skill in the art having benefit of the description herein.
DETAILED DESCRIPTION:
For the purpose of promoting an understanding of the principles of the invention, reference will now be made to the embodiment illustrated in the figures and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, such alterations and further modifications in the illustrated system, and such further applications of the principles of the invention as illustrated therein being contemplated as would
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normally occur to one skilled in the art to which the invention relates.
It will be understood by those skilled in the art that the foregoing general description and the following detailed description are exemplary and explanatory of the invention and are not intended to be restrictive thereof.
Reference throughout this specification to “an aspect”, “another aspect” or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrase “in an embodiment”, “in another embodiment” and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.
The terms "comprises", "comprising", or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a process or method that comprises a list of steps does not include only those steps but may include other steps not expressly listed or inherent to such process or method. Similarly, one or more devices or systems or elements or structures or components proceeded by "comprises... a" does not, without more constraints, preclude the existence of other devices or other systems or other elements or other structures or other components or additional devices or additional systems or additional elements or additional structures or additional components.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The system, methods, and examples provided herein are illustrative only and not intended to be limiting.
The terms “a” and “an” herein do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced items.
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The terms “having”, “comprising”, “including”, and variations thereof signify the presence of a component.
Now the present invention will be described below in detail with reference to the following embodiment.
Example 1
Materials
Pluronic F127 was purchased from “Upashna Enterprises” (Gurugram, India), Soya Lecithin was purchased from “Chembio lifesciences” (Ghaziabad, India), and other chemicals like Isopropyl myristate, ascorbic acid, parabens, disodium hydrogen phosphate, potassium dihydrogen phosphate, sodium chloride and vitamin E oil were also of “analytical grade”.
Methods
In the present study an attempt were made to formulate and evaluate topical organogel of insulin. 6 batch of formulation were prepared with varying concentration of soya lecithin (F1-F3) and pluronic F127 (F4-F6) and evaluated for various formulation parameters.
Example 2
Drug-excipients compatibility study Insulin with pluronic F 127 The FTIR spectrum of insulin with pluronic F127 was characterized by the main peak of insulin spectra was COO- which was observed at 1641.20 cm-1 and the main peak of pluronic F 127 spectra is O-H which is observed at 1351.18 cm-1, C-O-C bond observed at 1249 cm-1 and C-O stretching. The observed results from spectra show that insulin is compatible with pluronic F 127.
Table 1: FTIR spectrum of insulin with pluronic F127
Observed insulin Peak (cm-1)
Interpretation of chemical functional groups
1641.20
COO- stretching
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1351.18
O-H bond
Insulin with soya lecithin The FTIR spectrum of insulin with soya lecithin was characterized and the main peak of insulin COO- was observed at 1639.72 cm-1 and 1383.48 cm-1, while the main peaks of soya lecithin spectra were 1220.83 cm-1 indicating the presence of a C-O bond, and 1082.32 cm-1 with low intensity showing the existence of a C-C bond present in molecules. Further the results indicate that insulin is compatible with soya lecithin.
Table 2: FTIR spectrum of insulin with soya lecithin
Observed insulin Peak (cm-1)
Interpretation of chemical functional groups
1639.72
COO- stretching
1383.48
COO- stretching
Drug with polymers (mixture) The FTIR spectrum of mixture was characterized by the main peak of insulin spectra was COO- which was observed at 1644.68 cm-1 and 1374.39 cm-1. The main peaks of pluronic F 127 was observed at 2854.89 cm-1 C-H stretching vibration, 1251.59 cm-1 C-O-C bonds and 1109.46 cm-1 C-O stretching. The main peaks of soya lecithin were observed at 1421.17 cm-1 indicating the presence of C-O-H bond and 1143.20 cm-1 C-O bond present in molecule.From the spectra it is observed that there is no drug and polymer interaction were found and according to the result it is compatible.
Table 3: FTIR spectrum of insulin with polymers (mixture)
Example 3
Observed insulin Peak (cm-1)
Interpretation of chemical functional groups
1644.68
COO- stretching
1374.39
COO- stretching
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Physical parameter of organogel
The organogel formulation was observed for physical appearance and other organoleptic characteristics. All formulations were found to be odorless, homogeneous, washable, Off-white in color, greasy, and stable without any sign of phase separation (Table 4).
Table 4: Physical appearance and organgoleptic charachteristics
S. No.
Formulation
Code
Homogeneity
Smell
Colour
Gelation nature
Washabaility
Phase separation
1
F1
Homogeneous
Odorless
Off-white
Greasy
Washable
No phase separation
2
F2
Homogeneous
Odorless
Off-white
Greasy
Washable
No phase separation
3
F3
Homogeneous
Odorless
Off-white
Greasy
Washable
No phase separation
4
F4
Homogeneous
Odorless
Off-white
Greasy
Washable
No phase separation
5
F5
Homogeneous
Odorless
Off-white
Greasy
Washable
No phase separation
6
F6
Homogeneous
Odorless
Off-white
Greasy
Washable
No phase separation
The organogel formulation was observed for physical appearance and other organoleptic characteristics. All formulations were found to be odorless, homogeneous, washable, Off-white in color, greasy, and stable without any sign of phase separation (Table 4).
ii) Morphology characterization
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A light microscope examination of organogel at 10X has shown organogel as a bicontinuous system containing water molecules entrapped within oil (figure 4 (a)).
For the internal structure of TEM images of the organogel at 1.0µm clearly showed the gel as the microemulsion of the formulation (Figure 4 (b)).
iii) pH
The pH of all formulations was found to be in the range of 6 - 6.5 pH, which was in the normal range of skin pH and compatible with the skin. Therefore, all formulations are non-irritant to the skin (Table 5 and Figure 5).
iv) Spreadability
The spreadability of all formulations was found to be in the range of 0.35 g/cm to 0.52 g/cm. The value indicates that the organogel was easily spread with small shear, but an increase in the percentage of soya lecithin resulted in a decrease in the spreadability of the organogel. Oragnogel's spreadability decreases when polymer concentrations rise because of increased cross-linking between the polymers, which makes it harder for the gel to spread (Table 5 and Figure 5).
v) Viscosity
The viscosity of the organogel was found in the range of 2053 cps to 2816 cps for F1 to F3 and the range of 2522 cps to 3168 cps for F4 to F6. The occurrence of increased cross-linking in polymers with a rise in polymer concentration can be the cause of the corresponding increase in viscosity (Table 5 and Figure 5).
vi) Gel transition temperature
The gel transition temperature of the Organogel was found to be in the range of 35.3°C to 29°C for F1–F3 and the range of 36.3°C to 28°C for F4–F6. As the polymer concentration increased, the gel strength also increased, and the gelation occurred at a lower temperature (Table 5 and Figure 5).
Table 5: Characterization of organogel
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S. No.
Formulation
pH
Spreadability (g*cm/s)
Viscosity (cps)
Gelation temperature (°c)
1
F1
6.00 ± 0.02
0.35 ± 0.02
2058 ± 6.42
35.33 ± 1.15
2
F2
6.30 ± 0.02
0.38 ± 0.02
2561 ± 1.52
32.33 ± 1.52
3
F3
6.20 ± 0.02
0.51 ± 0.02
2816 ± 1.52
30.33 ± 2.51
4
F4
6.30 ± 0.03
0.55 ± 0.03
2522 ± 6.42
36.33 ± 1.52
5
F5
6.50 ± 0.01
0.45 ± 0.03
2723 ± 4.72
33.66 ± 4.72
6
F6
6.38 ± 0.02
0.52 ± 0.02
3168 ± 7.54
28.33 ± 0.57
Data presented as mean ± SD (n=3)
vii) Percentage drug content
The percentage drug content of organogel was found to be in the range of 79.11% to 96.43%. It was calculated by using the standard curve of insulin (Table 6).
Table 6: Percentage drug content of developed organogels
S. No.
formulation
Percentage drug content
1
F1
79.345 ± 0.5
2
F2
96.439 ± 0.2
3
F3
86.182 ± 0.1
4
F4
82.764 ± 0.1
5
F5
90.456 ± 0.5
6
F6
95.584 ± 0.1
Data presented as mean ± SD (n=3).
viii) In vitro drug release study
The drug release profile of the insulin topical organogel formulation was accomplished by diffusion cell. The outcomes of the in vitro release studies of all formulations are given below (Table 7).
Table 7: Percentage Cumulative drug release in 2 hours
S. No.
Formulation
% cumulative drug release
1
F1
78.51 ± 0.1
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2
F2
92.38 ± 0.5
3
F3
81.36 ± 0.1
4
F4
64.82 ± 0.8
5
F5
72.67 ± 0.9
6
F6
84.62 ± 0.1
Data presented as mean ± SD (n=3).
It was noted that soy lecithin concentration and pluronic concentration showed variation in the drug release profile.
Table 8: In vitro drug release of organogel formulations
Time (in min)
F1
(%)
F2
(%)
F3
(%)
F4
(%)
F5
(%)
F6
(%)
5
2.48± 0.05
3.11± 0.02
3.30± 0.03
3.76± 0.02
1.48± 0.01
2.22± 0.05
15
18.50± 0.09
19.29± 0.06
12.91± 0.05
11.75± 0.03
12.50± 0.05
19.29± 0.09
30
33.58± 0.03
36.56± 0.08
39.15± 0.01
37.19± 0.05
30.10± 0.06
43.56± 0.02
60
55.11± 0.06
62.38± 0.03
54.94± 0.08
52.53± 0.02
48.56± 0.03
62.38± 0.01
90
69.21± 0.05
83.62± 0.02
61.69± 0.04
61.06± 0.08
60.33± 0.05
69.32± 0.07
120
78.51± 0.03
92.38± 0.09
81.36± 0.05
64.82± 0.01
72.67± 0.08
84.62± 0.04
In formulations, F1–F3, the concentration of lecithin was gradually increased, and in formulations F4–F6, the concentration of pluronic was increased, resulting in a decrease in gel transition temperature, an increase in viscosity, and a gradual change in spreadability. The higher-viscosity formulations were much more stable and had better drug release. All
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formulations were fitted to a kinetic model belonging to first-order kinetics. However, after examining the parameter evaluation, it was found that the formulations F2 and F6 were better suited to the kinetic model and were consistent with the first-order and Higuchi models, which was a confirmation of the sustainability of the release system with matrix diffusion and drug delivery mechanisms that were based on the Super-Case II transport.
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We claim,
1. A insulin loaded topical Organogel formulation for the management of Diabetic wound comprising pluronic lecithin organogel and insulin.
2. The insulin loaded topical Organogel formulation as claimed in claim 1, wherein pluronic lecithin organogel comprises of Pluronic F127, Soy lecithin.
3. The insulin loaded topical Organogel formulation as claimed in claim 1, wherein Organogel formulation further comprises Isopropyl alcohol and myristic acid.
4. The insulin loaded topical Organogel formulation as claimed in claim 3, wherein Isopropyl alcohol and myristic acid integrate to form the ester known as “isopropyl myristate”.
5. The insulin loaded topical Organogel formulation as claimed in claim 3, wherein isopropyl myristate as a skin penetration enhancer.
6. The insulin loaded topical Organogel formulation as claimed in claim 1, wherein Organogel formulation is off-white, homogeneous, washable, and has a pH between 6 and 6.5.
7. The insulin loaded topical Organogel formulation as claimed in claim 1, wherein Organogel formulation has drug content range of 79.34 ± 0.5 – 96.43 ± 0.2 (%).
8. The insulin loaded topical Organogel formulation as claimed in claim 1, wherein Organogel formulation has (%) cumulative drug release range of 78.51 ± 0.1 – 92.38 ± 0.5 (%).
9. A process of preparation of insulin loaded topical Organogel formulation as claimed in claim 1 comprises the micro-emulsion method involves mixing the “aqueous and oil phases” at high shear.
Dated this 31/10/2023 G D Goenka University, Sohna Gurugram Road, Sohna, Haryana, India, 122103
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ABSTRACT
DoE Assisted Development of Insulin Loaded Topical Organogel Formulation for the Management of Diabetic Wound
The present invention relates to pharmaceutical science field, which aims at formulate and evaluate organogel of insulin that can serve as a topical administration for promoting enhanced wound healing in diabetic patients by providing sustained and localized delivery of drug to the wound site. The insulin organogel formulated by the micro-emulsion method involves mixing the “aqueous and oil phases” at high shear. The insulin loaded topical Organogel formulation for the management of Diabetic wound comprising pluronic lecithin organogel and insulin. The insulin loaded topical Organogel formulation has drug content range of 79.34 ± 0.5 – 96.43 ± 0.2 (%) and (%) cumulative drug release range of 78.51 ± 0.1 – 92.38 ± 0.5 (%). , Claims:We claim,
1. A insulin loaded topical Organogel formulation for the management of Diabetic wound comprising pluronic lecithin organogel and insulin.
2. The insulin loaded topical Organogel formulation as claimed in claim 1, wherein pluronic lecithin organogel comprises of Pluronic F127, Soy lecithin.
3. The insulin loaded topical Organogel formulation as claimed in claim 1, wherein Organogel formulation further comprises Isopropyl alcohol and myristic acid.
4. The insulin loaded topical Organogel formulation as claimed in claim 3, wherein Isopropyl alcohol and myristic acid integrate to form the ester known as “isopropyl myristate”.
5. The insulin loaded topical Organogel formulation as claimed in claim 3, wherein isopropyl myristate as a skin penetration enhancer.
6. The insulin loaded topical Organogel formulation as claimed in claim 1, wherein Organogel formulation is off-white, homogeneous, washable, and has a pH between 6 and 6.5.
7. The insulin loaded topical Organogel formulation as claimed in claim 1, wherein Organogel formulation has drug content range of 79.34 ± 0.5 – 96.43 ± 0.2 (%).
8. The insulin loaded topical Organogel formulation as claimed in claim 1, wherein Organogel formulation has (%) cumulative drug release range of 78.51 ± 0.1 – 92.38 ± 0.5 (%).
9. A process of preparation of insulin loaded topical Organogel formulation as claimed in claim 1 comprises the micro-emulsion method involves mixing the “aqueous and oil phases” at high shear.