FULLERENE-BIOTIN CONJUGATE FORMULATION AND PROCESS OF PREPARATION THEREOF

Abstract

ABSTRACT The present invention relates to a fullerene-biotin conjugate and formulations thereof for glucose metabolism and delivery of insulin to the cells. A derivative of fullerene polyethyleneimine fullerene is used as a carrier for biotin and a process of preparation thereof. Reference Fig 1 Dated this 12th Day of October 2024

Specification

Description:FIELD OF INVENTION
The present invention relates to enhanced drug delivery systems and nutraceuticals formulation comprising fullerene conjugated with biotin, designed for applications in managing blood glucose and peripheral neuropathy.

BACKGROUND OF THE INVENTION
Diabetes mellitus is a global health problem with a prevalence of 11.3% and is projected to rise continuously . According to the World Health Organization, 422 million people worldwide are suffering from diabetes and 1.5 million deaths are directly attributed to diabetes every year. Type 2 diabetes mellitus (T2DM) is the most common type of diabetes and usually results in macro- and micro-vascular complications and comorbidities in the body, such as diabetic retinopathy (DR), diabetic foot, and diabetic kidney disease (DKD) . These chronic complications have caused severe adverse impact on the quality of life of human beings. Therefore, the prevention and management of T2DM is of vital importance to alleviate its disease burden.
Dietary vitamin supplementation is an easy and economic strategy for both pre-diabetic and diabetic patients to help control glucose and lipid metabolism. Vitamins are generally classified as fat-soluble and water-soluble. Various studies have systematically evaluated the effects of fat-soluble vitamins in lowering the risk of T2DM and improving glycemic indices, such as vitamin A , vitamin D , vitamin E (and vitamin K . Specifically, vitamin D supplementation has been proved to significantly ameliorate status of fasting blood glucose (FBG), homeostatic model assessment of insulin resistance (HOMA-IR), and glycated hemoglobin (HbA1c) (10). Similarly, the role of water-soluble vitamins is also important in maintaining energy metabolism. For example, vitamin B1 (thiamine) is a co-enzyme for several enzymes, including transketolase which is essential in the non-oxidative branch of the pentose phosphate pathway (PPP). Many diabetic patients have been shown to have thiamine deficiency and lower blood thiamine pyrophosphate concentrations were associated with higher risk of DR . Moreover, vitamin B12 deficiency, usually caused by prolonged use of metformin, could result in severe oxidative stress and peripheral neuropathy. As frequent urination and excessive thirst are two of the main symptoms of T2DM patients, it is unknown whether water soluble vitamins would be lost along with the extra urine discharge, thereby exacerbate the water-soluble vitamin deficiency in T2DM patients. Therefore, the study of individual water-soluble vitamin supplementation is necessary to determine their blood status and effects on T2DM patients.

Biotin, or vitamin B7, is a water-soluble vitamin that acts as a prosthetic group of carboxylases. It serves as the essential cofactor for the 6 biotin-dependent carboxylases: acetyl-CoA carboxylase (ACC), geranyl-CoA carboxylase (GCC), 3-methylcrotonyl-CoA carboxylase (MCC), pyruvate carboxylase (PC), propionyl-CoA carboxylase (PCC), and urea carboxylase (UC) . The first physiological function of biotin was found in 1968 and proved to increase hepatic glucokinase transcription . Subsequently, many in vitro studies reported that biotin could stimulate pancreatic islet glucokinase activity and expression , increase insulin secretion and induce insulin receptor synthesis . Meanwhile, various in vivo studies have also proven the efficiency of biotin in ameliorating diabetic status. Lazo et al. investigated the effects of biotin in rodent pancreatic islets and confirmed that biotin supplementation could augment the proportion of beta cells and suppress mRNA expression of neural cell adhesion molecule. In animal studies, the results of glucose tolerance test (GTT) and insulin tolerance test (ITT) also supported the association between biotin treatment and improved tolerance condition.

Current researches have demonstrated that biotin deficiency could impair energy production by decreasing glucose utilization and oxidative phosphorylation. Chuahan et al. reported that biotin could regulate the glucokinase gene at the transcriptional stage in starved rats. In addition, insulin expression and secretion were found to be increased in response to biotin administration
Biotin is the water-soluble B vitamin that is an essential co-factor for 4-carboxylase enzymes each of which catalyzes an essential step in intermediary metabolism (Alternative Medicine Review, 2007). It is thought biotin improves abnormal glucose metabolism by stimulating glucose-induced insulin secretion in pancreatic ß-cells & by accelerating glycolysis in the liver and pancreas. Administration of high doses of biotin improved glycemic control (Furukawa Y, 1999).
Biotin is a water-soluble vitamin essential for all known organisms. Whereas animals cannot synthesize biotin, it is synthesized by microorganisms and plants and therefore widespread in the food supply at low concentrations relative to most water-soluble vitamins. Based on the available resources, biotin can be used for treating Hair and Nail Problems, Cradle Cap (Seborrheic Dermatitis), Diabetes, and Peripheral Neuropathy. However, the bioavailability and stability of biotin in the body are often limited by degradation during digestion.
Fullerene C60 is the primary allotrope of carbon and is a stable aggregate of 60 carbon atoms in the form of truncated icosahedron & is classified as nanomaterial (Kroto et al., 1991). It has been shown to induce inflammation, immunotoxicity & respiratory toxicity in rodents (Baker et al., 2008; Park et al., 2010). Many advancements in fullerene research have been made the first biological experiment involving C60 & other fullerene. Today, the conjugation of fullerene-based biomolecules has become a primary strategy for overcoming the intrinsic hydrophobic obstacles encountered by fullerene to utilize such molecules for biological purposes (Xinlin Yang et al., 2014). Since ROS has been shown to play a pivotal role in aging & various degenerative diseases fullerene conjugates with powerful antioxidative properties are promising candidates in the development of future innovative antioxidative against diabetes mellitus & associated diseases (Paradise WA et al., 2010).
Fullerenes, have been shown to possess excellent stability and biocompatibility, making them suitable carriers for bioactive compounds. By conjugating biotin with fullerene, it is possible to protect biotin from degradation, enhance its bioavailability, and provide sustained release, resulting in more effective blood glucose regulation.
OBJECTIVE OF THE INVENTION
The primary objective of the present invention is to develop a fullerene-biotin conjugated formulation.
It is another objective of the present invention to provide a fullerene-biotin conjugate that selectively targets biotin receptors, allowing for precise drug delivery.
Iti s another objective of the present invention to provide a unique carbon-based nanomaterial that provides high stability, large surface area, and unique electronic structure.
It is another objective of the present invention to develop formulations of fullerene-biotin for applications in managing blood glucose level as well as peripheral neuropathy.
It is another objective of the invention to provide a formulation with better pharmacological activity and reduced dosing frequency.
SUMMARY OF THE INVENTION
This summary is provided to disclose the concepts related to a fullerene-biotin conjugated formulation. This summary is not intended to identify essential features of the claimed subject matter nor is it intended for use in determining or limiting the scope of the claimed subject matter.
An embodiment of the invention relates to a fullerene-biotin conjugate formulation, comprising of:
- a fullerene (C60) derivative as a carrier,
- biotin as a vitamin supplement, and
- pharmaceutically acceptable inert excipients
to manage blood glucose levels
An aspect of the embodiment relates to a fullerene-biotin conjugate oral solid dosage form, comprising of:
- a polyethyleneimine fullerene (C60-PEI) in an amount of 0.25grams;
- biotin in an amount of 0.5 grams, and
- pharmaceutically acceptable inert excipients including but not limited to lubricants, fillers, binders, capsules, solvents, sweeteners, effervescing agents, natural flavourants or a combination thereof.
Another embodiment of the invention relates to A process for preparation of fullerene- biotin conjugate, comprising the steps of:
- Derivatising fullerene (C60) with ethylenediamine (EI) to form a polyethyleneimine fullerene( C60-PEI),
- Adding a mixture of biotin and ethylene-dichloride (EDC) to polyethyleneimine fullerene( C60-PEI) to form a resultant mixture,
- Dialyzing the resultant mixture to form the fullerene-biotin conjugate (C60-PEI-Biotin).

BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 is route of synthesis of the conjugate
Figure 2 shows a FTIR spectra of fullerene derivative (C60-PEI)
Figure 3 shows FTIR spectra of C60-PEI-Biotin conjugate
Figure 4 shows field emission scanning electron microscope of the conjugate
Figure 5 shows the particle size distribution by intensity and the polydispersity index of the conjugate.
Figure 6 1H NMR of the compound C60-PEI-Biotin
DETAILED DESCRIPTION OF THE INVENTION
Some embodiments of this disclosure, illustrating all its features, will now be discussed in detail. The words "comprising," "having," "containing," and "including," and other forms thereof, are intended to be equivalent in meaning and be open ended in that an item or items following any one of these words is not meant to be an exhaustive listing of such item or items, or meant to be limited to only the listed item or items.
Although any methods similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present disclosure, the exemplary methods are now described. The described embodiments are merely exemplary of the disclosure, which may be embodied in various forms.
Various modifications to the embodiment will be readily apparent to those skilled in the art and the generic principles herein may be applied to other embodiments. However, one of ordinary skill in the art will readily recognize that the present disclosure is not intended to be limited to the embodiments illustrated, but is to be accorded the widest scope consistent with the principles and features described herein.
The following sections of this article will provide various embodiments of the current invention with references to the accompanying drawings, whereby the reference numbers utilised in the picture correspond to like elements throughout the description. However, this invention is not limited to the embodiment described here and may be embodied in several other ways. Instead, the embodiment is included to ensure that this disclosure is extensive and complete and that individuals of ordinary skill in the art are properly informed of the extent of the invention. Numerical values and ranges are given for many parts of the implementations discussed in the following thorough discussion. These numbers and ranges are merely to be used as examples and are not meant to restrict the claims' applicability. A variety of materials are also recognised as fitting for certain aspects of the implementations. These materials should only be used as examples and are not meant to restrict the application of the innovation.
The term conjugate refers to a molecule formed by covalently bonding two distinct entities-in this case, biotin and fullerene. A conjugate combines the properties of both components, such as the metabolic role of biotin and the stability provided by fullerene.
The term Biotin-Fullerene Conjugate refers to a covalent bond between biotin and fullerene (C60), designed to improve the stability, bioavailability, and efficacy of biotin when administered as a health supplement. The conjugate ensures biotin is delivered effectively to the body, enhancing its impact on blood glucose regulation.
The term "Carrier Molecule" refers to a compound, such as fullerene, used to transport or enhance the delivery of an active ingredient (e.g., biotin) in pharmaceutical or health supplement formulations. The carrier improves the absorption, distribution, and stability of the active ingredient.
The term glucose metabolism refers to the biochemical process by which the body converts glucose (sugar) into energy. Proper regulation of glucose metabolism is essential for maintaining healthy blood sugar levels, particularly in individuals with metabolic disorders like diabetes.
The term a pharmaceutical formulation or composition refers to a mixture of an active ingredient (such as the biotin-fullerene conjugate) and excipients, formulated for administration to a subject for therapeutic, preventative, or health-promoting effects.
An embodiment of the invention relates to a fullerene-biotin conjugate formulation, comprising of:
- a fullerene (C60) derivative as a carrier,
- biotin as a vitamin supplement, and
- pharmaceutically acceptable inert excipients
The management of blood glucose levels is critical for individuals with metabolic disorders such as diabetes. Biotin, also known as Vitamin B7, is known to support the body's glucose metabolism by enhancing insulin sensitivity and promoting the efficient utilization of glucose. However, the bioavailability and stability of biotin in the body are often limited by degradation during digestion.
The present invention provides compositions comprising biotin-fullerene conjugates, where fullerene (C60) acts as a carrier for biotin (Vitamin B7), which plays a crucial role in glucose metabolism and insulin function. Biotin is covalently attached to functionalized fullerene through a linker, such as an amide bond, ensuring improved bioavailability and protection against degradation.
In accordance with an embodiment of the invention, Biotin is covalently bonded to the fullerene structure, forming a fullerene-biotin conjugate. Biotin acts as a targeting moiety, directing the conjugate to insulin receptor-expressing cells, such as those in the liver, muscle, and adipose tissues. Biotin's high affinity for biotin receptors allows for selective targeting and enhanced uptake of the fullerene-biotin conjugate by cells involved in glucose metabolism.
In a further aspect, biotin is used in an amount of 2-6 mM (milli moles) per dosage form in the composition.
In accordance with an embodiment of the invention, the fullerene used in this invention is selected from C60 or its derivatives, owing to its ability to carry biomolecules and protect them from degradation. Fullerenes have been extensively studied as nanocarriers due to their stable, cage-like structure and the ability to interact with a variety of therapeutic agents, including peptides and proteins. In this formulation, fullerene serves as the primary nanocarrier for insulin delivery.
In an aspect of the embodiment, a derivative of fullerene is used to form a conjugate- a polyethyleneimine fullerene (C60-PEI).
In an aspect of the embodiment, the C60- PEI is composed of ethylenediamine and fullerene C-60 compound.
In an aspect, ethylenediamine is used in an amount of 70-150 ml, and fullerene in an amount of 0.3-0.8 grams.
Synthesis of the conjugate
An embodiment of the invention relates to a process for synthesis of C60-PEI-biotin conjugate, comprising the steps of:
- Derivatising fullerene (C60) with ethylenediamine (EI) to form a polyethyleneimine fullerene( C60-PEI),
- Adding a mixture of biotin and ethylene-dichlroide (EDC) to polyethyleneimine fullerene( C60-PEI) to form a resultant mixture,
- Dialyzing the resultant mixture to form the fullerene-biotin conjugate (C60-PEI-Biotin).
Synthesis of a derivative of fullerene (C60-PEI)
Figure 1 encompasses the process of synthesis of the fullerene-biotin conjugate, comprising the steps of:
- dissolving ethylenediamine (H2NCH2CH2NH2 in an ethanol-water mixture (ethanol: water ¼ 5:1, 240 ml)
- adding the resultant mixture added drop-wise to a stirring dry toluene solution (500 ml) containing C60 and NaOH to form a mixture
- After stirring the mixture at room temperature under the protection of Aziridin for 7 days, the mixture was evaporated and dried in a vacuum at 60°C for 24 h, then purified by repeated rinsing with ethanol: water mixture (ethanol: water ¼ 6:1) and filtrations,
- the resulting solid products were dried in a vacuum at 60°C for 24 h. Finally, the above product (250 mg), concentrated HCl (50 ml), and aziridine (2.5 ml) were added into dichloromethane (150 ml), stirred, and refluxed at 40°C under the protection of Ar for 24 h. T
- The final product (C60-PEI) will be obtained by filtering and washing in order of CH2Cl2 10 times, methanol twice, and deionized water several times.

Synthesis of the conjugate
- Biotin (3.5 mM) and ethylene dichloride EDC will be added to a solution of C60-PEI in 10 mM phosphate temperature in the dark for 24 h, and
- finally dialyzed by a membrane for 3 days to remove free biotin and EDC.
- The resulting product (C60-PEI-Biotin) was dried in a vacuum at 40°C for 24
In an aspect of the embodiment, biotin is used in an amount of 2-6 mM, preferably 3-4mM.
In an aspect of the embodiment, ethylene dichloride is mixed with biotin in an amount of 3-8mM, preferably 4-6 mM.
An embodiment of the invention relates to oral solid dosage forms of the biotin-fullerene conjugate.
In an aspect of the embodiment, the formulation is in the form of a capsule, effervescent tablet, chewable tablets or a gummy.
In an aspect of the embodiment, the formulation comprises of the biotin-fullerene conjugate and pharmaceutically acceptable inert excipients.
In an aspect of the embodiment, the pharmaceutically acceptable inert excipients are selected from lubricants, fillers, binders, capsules, solvents, sweeteners, effervescing agents, natural flavourants or a combination thereof.
In accordance with an aspect of the embodiment, the formulation is in the form of an oral capsule comprising of fullerene-biotin conjugate, a filler, a lubricant and a gelatine capsule.
In an aspect of the embodiment, the oral capsule comprises of fillers such as but not limited to microcrystalline cellulose, lactose monohydrate, starch, dicalcium phosphate (DCP), magnesium stearate, silicon dioxide, calcium carbonate, mannitol, or cellulose derivatives.
In an aspect of the embodiment, the oral capsule comprises of a lubricant selected from magnesium stearate, stearic acid, talc, calcium stearate, glyceryl behenate or a combination thereof.
In an aspect of the embodiment, the biotin-fullerene conjugate is prepared by covalently bonding biotin to fullerene. the resulting conjugate is mixed with microcrystalline cellulose and magnesium stearate, and encapsulated in a gelatine capsule.
In an aspect of the embodiment, the formulation is in the form of effervescent tablets, comprising of the conjugate, effervescent agents, sweeteners, and natural flavouring agents.
In an aspect of the embodiment, effervescent tablets include sodium bicarbonate, citric acid, tartaric acid, malic acid, sodium carbonate, potassium bicarbonate, ascorbic acid, fumaric acid, ascorbic acid, fumaric acid or a combination thereof.
In an aspect of the embodiment, the effervescent tablets include sweeteners selected from natural or artificial sweeteners such as aspartame, sorbitol, xylol, sucralose steviol or other such similar sweeteners.
In an aspect of the embodiment, the effervescent tablets include flavouring agents selected from natural or artificial fruit flavours, vanilla, mint, cinnamon, berry or the like.
An aspect of the embodiment, relates to a formulation in the form of chewable tablets comprising of the biotin-fullerene conjugate, a binding agent , a filler and a sweetener.
In an aspect of the embodiment, the chewable tablets contain a binding agent- pectin, a filler selected from the list given above, and a sweetener selected from the list given above.
An aspect of the embodiment, relation to a formulation in the form of a gummy, wherein the chewable gummy comprises of the fullerene-biotin conjugate, a gelling agent/ polymer, a sweetener, a coating agent and a flavourant.
In an aspect of the embodiment, the gelling agent used in the gummy is gelatine, the coating agent is beewswax and the sweetener and a flavouring agent is one selected from the list given above.
In an aspect of the embodiment, the fullerene- biotin conjugate is used in the oral solid dosage form in an amount of 0.5-5% by weight of the composition, comprising of biotin equivalent to 0.5-2% by weight of the conjugate.
In an aspect of the embodiment, a 500 milligrams of unit dosage form consists of 2.5-10 milligrams of the fullerene-biotin conjugate, comprising of biotin equivalent to 0.1-1 milligrams.
The present invention relates to compositions comprising fullerene- biotin conjugates, to provide a drug delivery system for biotin, resulting in enhanced drug delivery, absorption and better bioavailability of biotin. The present invention provides helps in maintaining the proper supply of insulin to the cells and has increased pharmacological activity.
EXAMPLES
The invention is illustrated by the following examples which are only meant to illustrate the invention and not to act as limitations. All embodiments apparent to a process there in the art are deemed to fall within the scope of the present invention.
Example 1: Synthesis of fullerene-biotin conjugate.
1. SYNTHESIS OF C60-PEI
In total, ethylenediamine (H2NCH2CH2NH2, 100 ml) was dissolved in an ethanol-water mixture (ethanol: water ¼ 5:1, 240 ml) which was added drop-wise to a stirring dry toluene solution (500 ml) containing C60 (500 mg) and NaOH (10.0 g). After stirring at room temperature under the protection of Ar for 7 days, the mixture was evaporated and dried in a vacuum at 60°C for 24 h, then purified by repeated rinsing with ethanol: water mixture (ethanol: water ¼ 6:1) and filtrations.
The resulting solid products were dried in a vacuum at 60°C for 24 h. Finally, the above product (250 mg), concentrated HCl (50 ml), and aziridine (2.5 ml) were added into dichloromethane (150 ml), stirred, and refluxed at 40°C under the protection of Ar for 24 h. The final product (C60-PEI) will be obtained by filtering and washing in order of CH2Cl2 10 times, methanol twice, and deionized water several times. The sequence of steps involved in synthesis is summarized in Figure 1
2. CONJUGATION OF BIOTIN TO C60-PEI
Biotin (3.5 mM) and EDC (5 mM) will be added to a solution of C60-PEI in 10 mM phosphate temperature in the dark for 24 h, and finally dialyzed by a membrane (MWCO ¼ 10,000, Spectrum Laboratories Inc) for 3 days to remove free biotin and EDC. The resulting product (C60-PEI-Biotin) will be dried in a vacuum at 40°C for 24 h.

Example 2 Characterization of the structure with spectroscopy
Structural characterization of the synthesized compound was done by instrumental methods of analysis like IR, 1H NMR, MS and Zeta-Sizer
Spectral analysis of all the synthesized derivatives was carried out using Perkin-Elmer Spectrum 400 FTIR for IR spectra, Bruker Avance II for 1H NMR, Waters Alliance 2795, Q-TOF Micromass Mass spectrometer for MS and , and Malvern analytical zeta sizer Nano ZS for zeta size.
IR SPECTROSCOPY
The peaks in the IR spectrum give an idea about the probable structure of the compound's IR region ranges between 4000-600 cm-1. Quanta of radiation from this region of the spectrum corresponds to energy differences between different vibrational levels of the molecule. IR spectra were recorded on a Perkin-Elmer FT-IR spectrometer (Sharma YR., 2013).

FTIR results showed three bands that graft the PEI to C60. This was confirmed by the strong C-N (~1095 cm-1) vibrations, N-H (~3387.5 cm-1) vibrations, and C-H (2922 cm-1) vibration of PEI due to C60 molecules shown in figure 2 and interpretation in table 1. The biotin molecule was reacted to form amide linkage to the termini of branched PEI. The FTIR result showed the biotin conjugated to C60-PEI was confirmed by strong C-N (~1310 cm-1) vibrations, N-H (~3307 cm-1) vibrations, C-H (~2857 cm-1) vibrations and O-H (2931cm-1) vibrations shown in figure 3 and interpretation in table 2.
Table 1: Interpretation of C60-PEI

S.No. IR Absorptions Band (cm-1) Interpretation
1. 3388.7 N-H stretching of amides
2. 1095 C-N stretching of amines
3. 2922 C-H stretching of alkane

Table 2: Interpretation of C60-PEI-Biotin

S.No. IR Absorptions Band (cm-1) Interpretation
1. 3307 N-H stretching of amides
2. 2857 C-H stretching of alkane
3. 1310 C-N stretching of amine
4, 2931.5 O-H stretching of carboxylic acid

1H NMR SPECTROSCOPY
1H NMR spectroscopy enables us to record differences in magnetic properties of the various magnetic properties of the various magnetic nuclei present and to deduce how many different kinds of environmental are in molecules and also which atoms are present in neighboring groups. The 1H NMR spectra, enable us to know different magnetic environment corresponding to protons in molecules. 1H NMR was recorded on Bruker Avance II 400 spectrophotometer operating at 400.MHz. data present in chemical shift-values (d) (Chatwal GR et al., 2019).
1H NMR of C60-PEI (CDCl3): d 2.1761 (H, NH, NH2). A single peak was observed as the protons of the polyethylene chain exist in a similar environment (figure 6).
Zeta Potential
Zeta potential measurements are relevant only for samples with size in the sub 5 µm region. If the sedimentation or significant aggregation of the sample occurs before or during the measurement the system is not suitable for zeta measurement since nanoparticle electrophoretic mobility is strongly compromised. Zeta potential was recorded on Malvern analytical zeta sizer Nano ZS (Standard Guide for Zeta Potential, 2012). The particle size of the fullerene-biotin conjugate was 1197 nm with the PDI value of 0.721. (Figure 5)

Field Eimmison Scanning Electron Microscope
A powdered sample of the conjugate was settled on the slide, the sample was examined by the Hitachi SU-8010 field emission scanning electron microscope. There was the presence of crystals and rod-like structures which were entrapped in the hollow spheres within the range of 500 nm to 5 µm shown in Figure 4
Structural characterization of the synthesized compounds was done by instrumental methods like FT-IR, 1H NMR, Zeta size, and MS. FT-IR showed three bonds due to grafting of PEI to C60 i.e., 1095 cm-1, 3387.5 cm-1, & 2922 cm-1 corresponds to C-N, N-H & C-H vibrations of PEI due to C60 molecules. The conjugation of biotin with C60-PEI was confirmed by the frequencies 1310 cm-1, 3307 cm-1, 2287 cm-1, and 2931 cm-1 correspond to the different vibrations i.e., C-N, N-H, C-H and O-H. There was the presence of crystals and rod-like structures shown in the hollow sphere or fullerene within the range of 500 nm to 5 µm. the size of the compound C60-PEI-Biotin was 1197 nm with a PDI value of 0.721. 1H NMR peak of C60-PEI (CDCl3): d 2.1761 (H, NH, NH2). There was a single peak observed as the protons of the polyethylene chain.

Example 3 Oral Capsules of Fullerene- Biotin conjugate
Oral Capsules
Table 3:
Biotin-Fullerene Conjugate: 10 mg (equivalent to 1 mg of biotin)
Microcrystalline Cellulose: 100 mg (filler)
Magnesium Stearate: 2 mg (lubricant)
Gelatin Capsule (Size 0): 1 capsule
The biotin-fullerene conjugate was prepared by covalently bonding biotin to fullerene as shown in example 1. The resulting conjugate was mixed with microcrystalline cellulose and magnesium stearate, and encapsulated in a gelatin capsule.
Example 4 Effervescent Tablets of conjugate
Table 4:
• Biotin-Fullerene Conjugate: 10 mg (equivalent to 1 mg of biotin)
• Sodium Bicarbonate: 150 mg
• Citric Acid: 150 mg
• Sorbitol: 50 mg
• Natural Flavoring: 5 mg
• Stevia: 2 mg
The conjugate was prepared as shown in Example 1 and mixed with effervescent agents (sodium bicarbonate and citric acid), sweeteners, and natural flavors to form effervescent tablets.
Example 5 Chewable Tablets
Table 5:
• Biotin-Fullerene Conjugate: 5 mg (equivalent to 0.5 mg of biotin)
• Xylitol: 300 mg
• Pectin: 50 mg
• Flavoring: 10 mg
• Calcium Stearate: 2 mg
The conjugate as shown in example 1 was combined with pectin, xylitol, and natural flavors, and compressed into chewable tablets.
Example 6: Gummies
Table 6
• Biotin-Fullerene Conjugate: 2.5 mg (equivalent to 0.25 mg of biotin)
• Gelatin: 300 mg
• Glucose Syrup: 150 mg
• Natural Fruit Extract: 50 mg
• Beeswax: 10 mg
The conjugate as shown in example 1 was mixed with gelatin and glucose syrup to form gummies, flavored with fruit extracts and coated with beeswax.
, Claims:CLAIMS:
We claim:

1. A fullerene-biotin conjugate formulation, comprising of:
- a fullerene (C60) derivative as a carrier,
- biotin as a vitamin supplement, and
- pharmaceutically acceptable inert excipients to manage blood glucose levels.
2. The fullerene-biotin conjugate formulation as claimed in claim 1, wherein the fullerene (C60) derivative is a polyethyleneimine fullerene (C60-PEI).
3. The fullerene-biotin conjugate formulation as claimed in claim 2, wherein polyethyleneimine fullerene consists of ethylenediamine in an amount of 70-150 ml, and fullerene in an amount of 0.3-0.8 grams.
4. The fullerene-biotin conjugate formulation as claimed in claim 1, wherein biotin is present in an amount of 2-6 mM.
5. The fullerene-biotin conjugate formulation as claimed in claim 1, wherein the formulation is in the form of an oral solid dosage formulation comprising of the fullerene-biotin conjugate in an amount of 0.5-50% by weight of the formulation.
6. The fullerene-biotin conjugate formulation as claimed in claim 5, wherein the formulation is in the form of a capsule, effervescent tablet, chewable tablets or a gummy.
7. The formulation as claimed in claim 1, wherein the pharmaceutically acceptable inert excipients include lubricants, fillers, binders, capsules, solvents, sweeteners, effervescing agents, natural flavourants or a combination thereof.
8. A fullerene-biotin conjugate oral solid dosage form, comprising of:
- a polyethyleneimine fullerene (C60-PEI) in an amount of 0.25grams;
- biotin in an amount of 0.5 grams, and
- pharmaceutically acceptable inert excipients including but not limited to lubricants, fillers, binders, capsules, solvents, sweeteners, effervescing agents, natural flavourants or a combination thereof.
9. A process for preparation of fullerene- biotin conjugate, comprising the steps of:
- Derivatising fullerene (C60) with ethylenediamine (EI) to form a polyethyleneimine fullerene( C60-PEI),
- Adding a mixture of biotin and ethylene-dichlroide (EDC) to polyethyleneimine fullerene(C60-PEI) to form a resultant mixture,
- Dialyzing the resultant mixture to form the fullerene-biotin conjugate (C60-PEI-Biotin).
10. The process as claimed in claim 8, wherein 70-150 ml of ethylene diamine, and 0.3-0.8 grams of fullerene is used to form 200-300 milligrams of polyethyleneimine fullerene (C60-PEI).

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