AU2020103238A4

AU2020103238A4 - A formulation for treatment of diabetic complications and method of preparation thereof

Info

Publication number: AU2020103238A4
Application number: AU2020103238A
Authority: AU
Inventors: Neeraj Umesh Dwivedi; Pramod Rambhau Padole
Original assignee: Padole Pramod Rambhau Dr
Current assignee: Padole Pramod Rambhau Dr
Priority date: 2020-11-04
Filing date: 2020-11-04
Publication date: 2021-01-14
Anticipated expiration: 2028-11-04

Abstract

The present invention relates to a diabetic wound healing formulation and method of preparation thereof. The proposed composition has antiseptic, anti-inflammatory and antibacterial properties and can cure the injuries ulcers, bums, initial stage of Gangrene, wounds of the diabetic patient without causing any side effects. More particularly the present invention also has powerful moisturizing, protecting and healing ability to the skin. The proposed composition comprises of; Azdirachtaindica and SyzygiumCuminiO.01% Calcinated Magnesium w/v, 0.01% Silver nanoparticle w/w , 0.1% activated charcoal w/w, pure clarified butter of Buffalo, deionized water and 0.3% Ascorbic acid w/v respectively. Following invention is described in detail with the help of Figure 1 of sheet 1 showing oil proof butter paper, Figure 2 of sheet 1 showing cotton thread dressing gauze and Figure 3 of sheet 1 shows the strip. 1/2 100 101 102 Figure 1 103 Figure 2

Description

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A FORMULATION FOR TREATMENT OF DIABETIC COMPLICATIONS AND METHOD OF PREPARATION THEREOF

Technical field of invention:

Present invention in general relates to the field of pharmaceuticals and more particularly to diabetic wound healing formulation and method of preparation thereof which can cure the injuries ulcers, burns, initial stage of gangrene, and wounds of the diabetic patient without causing any side effects.

Background of the invention:

Wound healing is a complicated and dynamic process, which involves a series of continuous, co-ordinated and sometimes overlapping of events including haemostasis, inflammation, proliferation, epithelization, maturation and remodeling of scar tissue. Despite the fact that the process of repair begin immediately after an injury in all tissues that all wounds go through similar phases of healing, specialized tissues such as liver, skeletal tissue and eye have distinctive forms of regeneration and repair and follow separate pathway and under chronic conditions they are further complicated.

When a person has diabetes, wounds can take longer to heal, which can increase the risk of infections and other complications developing. According to the Centers for Disease Control (CDC), around 30.3 million people in the United States have a type of diabetes, and many of these people experience complications caused by infected wounds. Minor wounds, cuts, and bums are an unfortunate but unavoidable part of life. However, for people with diabetes, these injuries can lead to serious health issues. Many people with diabetes develop wounds that are slow to heal, do not heal well, or never heal. Sometimes, an infection might develop. An infection can spread to tissue and bone near the wound or more distant areas of the body. In some cases, and without emergency care, an infection can be life-threatening or may even be fatal. Even when an infection does not develop in a wound, slow healing can adversely affect a person's overall health and quality of life.

There are lots of medicines including herbal and allopathic which are being used to control sugar level of blood in diabetic patients. The few medicines are invented for treating ulcer and wounds, burns and the initial stage of the Gangrene but sometimes these medicines cause infection and side effects in the body and are costly for the people. While discussing about the existing medicines for treating diabetic wounds, bums, initial stage of the gangrene and the ulcers there are not complete curing medicine. Some of these medicines never heal complications properly related to the diabetics. Some of them adversely affect to the patients. The medicines available nowdays makes more serious to wounds, ulcers, bums and the initial stage of the Gangrene. Sometime patients complain pus formation in the infected site. The existing medicines are individually use on any complication such as ulcers, burns, wounds and for the initial stage of the gangrene.

Although various attempts are made before, for providing various formulations for wound healing and few of them are discussed as- 201841048015 discloses diab-wound-ayur-heal-an ayurvedic wound healing formulation for diabetic non healing ulcer-wound, vericos ulcers, pressure ulcers, bed sores, burn injuries and highly potential in treating and preventing the above said clinical conditions, EP2114386 discloses pharmaceutical composition for treatment of diabetic complications, IN5076/CHE/2015 discloses "diawound care" ayurvedic wound healing formulation for diabetic non healing ulcer and a process for preparation of the same, CN104189906 discloses pharmaceutical composition for treatment of diabetic complications, EP2916876A1 discloses methods and compositions for wound healing. The '0 invention relates to methods and compositions for wound healing. EP2224815A1 discloses silver nanoparticles with specific surface area and a method for producing them.

The above mentioned formulations for wound healing suffer various drawbacks therefore to overcome the related issues there is need to introduce an improved and efficient composition for the same. Hence the present invention provides a diabetic wound healing formulation and method of preparation thereof.

Object of the invention:

Primary object of the present invention is to provide a diabetic wound healing formulation and method of preparation thereof.

Another object of the present invention is to cure multi targeted complication like injuries, ulcers, bums, initial stage of gangrene, and wounds of the diabetic patient without causing any side effects.

Yet another object of the present invention is to provide healing ability, moisturizing property, anti-inflammatory, antiaging and antiseptic property.

Yet another object of the present invention is to increase improve completely curiosity ability, no any side effects, less manufacturing cost and absolutely safe to use for the complications of the diabetic patients.

Other objects, features and advantages will become apparent from detail description and appended claims to those skilled in art.

Summary of the invention:

Accordingly following invention provides a diabetic wound healing formulation and method of preparation thereof which cure multi targeted complication like injuries, ulcers, burns, initial stage of gangrene, and wounds of the diabetic patient without causing any side effects. '0 The proposed composition comprises of; Azadirachta Indica and Syzygium Cumini stem bark 0.01% Calcinated Magnesium w/v, 0.01% Silver nanoparticle w/w , 0.1% activated charcoal w/w, pure clarified butter of Buffalo, deionized water and 0.3% Ascorbic acid w/v respectively. All these constituents play vital role while dealing with biological functions in the cells such as cell repair, free radical stabilization, accelerating healing process, balancing hormones, enzymes, minerals, lipids and glucose inside the cell if it is diabetes type2, protecting cells from oxidative damage, protection from toxins releasing by the bacteria.

Brief description of drawing: This invention is described by way of example with reference to the following drawing where,

Figure 1 of sheet 1 shows oil proof butter paper Where, 100 denotes the smooth layer of the oil proof white butter paper

101 denotes breadth of the oil proof white butter paper 102 denotes length of the oil proof white butter paper

Figure 2 of sheet 2 shows cotton thread dressing gauze Where, 103 denotes cage form cotton thread dressing gauze

Figure 3 of sheet 2 shows the strip Where, 104 denotes breadth of the strip 105 denotes length of the strip

In order that the manner in which the above-cited and other advantages and objects of the invention are obtained, a more particular description of the invention briefly described above will be referred, which are illustrated in the appended drawing. Understanding that these drawing depict only typical embodiment of the invention and therefore not to be considered limiting on its scope, the invention will be described with additional specificity and details through the use of the accompanying drawing.

'0 Detailed description of the invention:

The present invention relates to a diabetic wound healing formulation and method of preparation thereof which cure multi targeted complication like injuries, ulcers, burns, initial stage of gangrene, and wounds of the diabetic patient without causing any side effects. In proposed invention the medicine has been formulated from antidiabetic plants stem bark plants names such as Azadirachta Indica stem bark and Syzygium Cumini stem bark and its freshly extracted liquid 8 ml of stem bark then 0.01% Calcinated Magnesium w/v, 0.01% Silver nanoparticle w/w, 0.1% activated charcoal w/w, pure clarified butter of Buffalo, deionized water and 0.3% Ascorbic acid w/v respectively.

In present invention primarily the phytochemical analysis of Azadirachta Indica were done in vivo using the methanolic extract. All the constituents determined by the phytochemical analysis. Then all the secondary metabolites were also determined present inside the extract of the Azadirachta Indica stem bark. Same procedures were repeated for the phytochemical analysis of the stem bark of the Syzygium Cumini methanolic extract. Consequently all the prescribed constituents were determined. Wherein the formulation involves following tests and analysis

Phytochemical Analysis A) Syzygium Cumini

S.no Phytoconstituents Aqueous Methanol Ethanol 1 Saponin ++ ++

2 Tannin ++ +++ +++ 3 Flavonoid + ++ ++ 4 Terpenoids ++ ++

+ 5 Alkaloid + ++ ++ 6 Phenols + ++

+ 7 Carbohydrate + ++ ++

8 Glycosides + +++ +++

B) Azadirachta Indica

S.No Phytoconstituents Methanol Chloroform 1 Alkaloids ++ 2 Glycosides ++

3 Saponins +++

4 Steroid ++ ++

5 Flavonoids ++

6 Tannins +++ ++

7 Phenols ++ +

Antioxidant Test The antioxidants are the substance that inhibits the oxidation. Oxidation is a chemical reaction in which the free radicals are generated which has unpaired electrons thereby leading the chain reaction that may damage the cells of organism. Antioxidant test were done on the way to determine the radical scavenging activity of the methanolic extract of Syzygium

Cumini plants stem bark and the same test were performed for Azadirachta Indica stem bark extract. The absorbance were measured at 517nm using instrument UV-Spectrophotometer at various concentrations and on the basis of data obtained the radical scavenging activity were shown in the extract of the stem bark. On the basis of absorbance obtained from antioxidant test it becomes easier to determine inhibition concentration value of stem bark of both the plants mentioned above i.e.IC50 value. And on the basis of IC50 Value the effective concentration i.e EC50 value. Were also determined which shows effectiveness of the constituents present inside the extract of the stem bark. The Antioxidant test performed for Syzygium Cumini and Azadirachta Indica such as;

a) Antioxidant test of Syzygium Cumini; The absorbance at 517nm wavelength were measured with standard 0.004% DPPH for the antioxidant activity of the Syzygium Cumini bark. And percentage of the radical scavenging activity were determined on the basis of the control ascorbic acid and the absorbance data obtained.

Sr.No. Sample ID Type Concentration Absorbance %RSA 1 Blank 01 Standard 0.000 0.076 2 S.Cumini Standard 0.250 0.128 58.97% 3 S.Cumini Standard 0.500 0.162 64.23% 4 S.Cumini Standard 0.750 0.197 66.55% 5 S.Cumini Standard 1.000 0.274 67.54%

b) Antioxidant Test of Azadirachta Indica; The absorbance at 517nm wavelength were measured with standard 0.004% DPPH for the antioxidant activity of the Azadirachta Indica bark. And percentage of the radical scavenging activity were determined on the basis of the control ascorbic acid and the absorbance data obtained.

Sr.No. Sample ID Type Concentration Absorbance %RSA 1 Blank 01 Standard 0.000 0.077 2 A.Indica Standard 0.250 0.131 58.01% 3 A.Indica Standard 0.500 0.167 63.13% 4 A.Indica Standard 0.750 0.207 64.85% 5 A.Indica Standard 1.000 0.252 69.71%

Antidiabetic test

The antidiabetic test of both the plants extract were done using alpha Amylase and successfully it determines that both the plants are antidiabetic in nature.

Antibacterial Test Mainly three bacteria which are very much harmful to the skin of humans having diabetes and the bacteria's responsible for the infection were isolated named as Staphylococcus aureus, Escharichia Coli and Pseudomonas aerguinosa. Out of these S.aures and Pseudomonas are highly toxic to the skin and the tissue of human body. The methanolic extract of the plants stem bark were used to know its antibacterial activity against S.aureus, E.Coli and Pseudomonas aeruginosa.

XRD Analysis The spectrums of all the inorganic and organic constituents induced into the formulation were determined by the XRD method, the peaks obtained from the XRD instrument were shown string elemental signal.

Herein the present formulation consists of antioxidants which tackle with free radicals and terminate its chain. This formulation also contains Vitamin C, Calcinated magnesium and the minerals which requires in the cell repair. Silver nanoparicles were added which plays active '0 role in the wound healing which is attributed by silver and along with its distinctive role in preventing infection this nanoparticle also drive in differentiation of fibroblasts into myo fibroblast, which in turn promotes the wound contraction and fasten healing rat., activated Charcoal added which absorbs toxic toxins from the tissues, two plants stem bark used in it plants named as Syzygium Cumini and Azadirachta Indica respectively. The effects of these constituents were determined doing clinical studies and hence present formulation is best multirole medicine among all the medicine present for the treatment.

Steps involved in preparing the proposed formulation:

The protocol of preparation is for one kg preparation. * Firstly take 800gms of wet stem bark of Syzygium Cumini plant and 200gms of wet bark of the Azadirachta Indica plants stem bark.

* Remove the peels present in outer surface and then grind both the stem bark separately in grinding machine for 5 minutes two times and remove chaff by using Sieve. * Make separately each in deionised water 0.01% Calcinated Magnesium w/v , 0.01% Silver nanoparticle w/w, 0.1% Activated Charcoal w/w, 0.3% Ascorbic acid w/v. * Mix both the grinded bark and add 6.5ml 0.01% Silver nanoparticle again and mix it well. * Take 300ml Buffalo pure clarified butter in iron Cauldron and maintain heat between 60 65 0C temperature then introduce mixture of bark into it. Now stir them. Then add 5ml 0.01% calcinated magnesium stir again, now add 6.5ml 0.01% Silver nanoparticle again stir, then add 4ml 0.1% activated charcoal stir it, and in last 0.100l of ascorbic acid. * Low the temperature at 360C and then make strip varying its different variety of strip According to the size of the wound, For small size strip; 3cm length, 2cm breadth and 4mm height, For medium size strip; 4cm length, 2cm breadth and 4mm height, For large size strip; 5cm length, 4cm breadth and 4mm height.

* Take extracted stem bark liquid by squeezing out bark of Syzygium Cumini plant and spray 1/2ml once it on each strip and cover the each strip with cotton thread dressing gauze which would be in distant aperture like sieve.

'0 e Pack the strip and place it in refrigerator. Before applying this strip on wounds, burns, and the gangrene firstly gently heat on any metal plate and immediately apply it on wound and before applying this strip on ulcers firstly surgically remove pus and the skin of ulcer site and gently heat on any metal plate and immediately apply it on surgical site of ulcer. Use 4-4cm oil proof butter paper on the strip and do dressing on it. * This strip would be applied onto the complications amide day and night.The complete recovery would be done during 29 to 33 days.

In the preferred embodiment of the invention regular practice of this medicine initiates healing and repair process in different phases as;

In phase 1 and 2 i.e Coagulation and Inflammation:

After the injury, a fibrin plug is form and the inflammatory cells are quickly recruited to the wounds. Coagulation is needed for homeostasis and for the wound protection. The fibrin consists of platelets which are embedded in a mesh work of polymerized fibrinogen. But in case of diabetic the fibrinogen such asfibronectin, vitronectin and thrombospondin, its action is to provide protection from bacteria but in diabetic patients it failed to provide protection.

Immediately after injury and burn the wound is hypoxic because of the blood vessels get damaged. When the present formulation is given the deleterious situation has beneficial effects and might prepare for next phase of healing. The process in diabetic patients are delayed this may take 3-4 days. The monocytes circulating take up residues at the injured site. As tissue macro phases, the fibroblasts and endothelial cells they form early granulation tissue that begins the process of contraction.

In phase 3 and 4 i.e Migration proliferation and remodeling:

After toned down inflammatory phase of the wound healing. The wound contraction begins. The balance between the contraction & keratinocyte closure has much to do on location of the wound and presence of complication. Present formulation prevents excessive deposition of some matrix proteins such as fibronectin and collagen. This happen in 11-15 days because '0 of sensory deficts. The diabetic patient have not protective symptoms guarding against heat and pressure. The infection can rapidly spread in diabetic ulcers. the limb -threatening cellulitis, abscesses osteomyelitis.

The continuous use of medicine helps remodeling in the tissues during the time period of 18 23 days and debridement starts which involves removal of the necrotic wound and callus. Thus remodeling or turnover of matrix might be inadequate which ultimately affects cell migration and stabilize the healed wound.

The present invention clinically studied on animals in which an evaluation will be conducted using 18 mice divided into three groups each group having six mice. Group 1- diabetic mice+ with wound, Group 2- control+ wound, Group 3- diabetic mice+ wound+ formulated medicine.

During this animal study on wistar mice 18 mice were selected for use and the individual weight of mice were between 250 to 300gms. The base line blood glucose of the all animal was determined normal. For the diabetes induction streptozotocin (STZ) was freshly dissolved in 0.1 M CITRATE BUFFER pH 4.5 the dose of STZ was administered by intra peritoneal (IP) injection of the 150 mg per kg to body weight for the IP injection then the mice was hold in one hand in dorsal position. The injection site was swabbed using ethanol and IP injected to caudal abdominal cavity using sterile needle. The diabetes was developed gradually and was assessed after 4 days .0.3cm incision wound was made using biopsy punch before making wound the anesthesia was given to the animals and the conditions was made in controlled for animal study. The medicine was applied on diabetic mice wound. The study was carried out till three weeks and successfully the wound healing activities occurs from this medicine and were recovered soon as compared to the control. Same study was done for the bum on mice.

Additional advantages and modification will readily occur to those skilled in art. Therefore, the invention in its broader aspect is not limited to specific details and representative embodiments shown and described herein. Accordingly various modifications may be made without departing from the spirit or scope of the general invention concept as defined by the appended claims and their equivalents.

Editorial Note 2020103238 There is 1 page of Claims only.

Claims

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:

1. A formulation for treatment of diabetic complications, comprises of Azadirachta indica stem bark, Syzygium cumini stem bark (8 ml extracted liquid), 0.01% Calcinated Magnesium w/v, 0.01% Silver nanoparticle w/w, 0.1% activated charcoal w/w, pure clarified butter of Buffalo, unionized water and 0.3% Ascorbic acid w/v.

2. The process for preparation of formulation for treatment of diabetic complications consisting of following steps; a. Firstly take 800gms of wet stem bark of Syzygium Cumini plant and 200gms of wet bark of the Azadirachta Indica plants stem bark; b. Remove the peels present in outer surface and then grind both the stem bark separately in grinding machine for 5 minutes two times and remove chaff by using Sieve; c. Mix both the grinded bark and add 6.5ml 0.01% Silver nanoparticle again and mix it well; d. Make separately each in deionised water 0.01% Calcinated Magnesium w/v , 0.01% Silver nanoparticle w/w, 0.1% Activated Charcoal w/w, 0. 3 % Ascorbic acid w/v; e. Take 300ml Buffalo pure clarified butter in iron Cauldron and maintain heat between 60-65 0C temperature then introduce mixture of bark into it; f. Then add 5ml 0.01% calcinated magnesium stir again, now add 6.5ml 0.01% Silver nanoparticle again stir, then add 4ml 0.1% activated charcoal stir it, and in last 0.100pl of ascorbic acid. g. Low the temperature at 360 C and then make strip varying its different variety of strip According to the size of the wound, h. Take extracted stem bark liquid of Syzygium Cumini plant and spray 1/2ml once it on each strip and cover the each strip with cotton thread dressing gauze which would be in distant aperture like sieve.