A synergistic process to enhance the bioavailability and bio efficacy of bioactive molecules

Abstract

The present invention discloses to enhance the bioavailability and bio efficacy of bioactive molecules. The process (100) comprises steps of isolating the bioactive molecules (101), high throughput screening of bioactive molecules (102), identification of synergistic molecules (103), precision particle engineering of bioactive molecules and synergistic molecules (104), uniform mixing of bioactive molecules and synergistic molecules (105), homogenization of the mixture (106) and spray drying of the homogenized mixture (107). The process is compatible with bioactive active molecules exhibiting a wide range of solubility. The process results in enhanced solubility hence facilitating increased bioavailability and bio efficacy.

Description

Internal Ref: OR26C001PCT01TITLE OF THE INVENTIONA synergistic process to enhance the bioavailability and bio efficacy of bioactive moleculesPriority Claim:

[0001] This application claims priority from the provisional application numbered 202441050510 filed with Indian Patent Office, Chennai on 2ndJanuary 2025 entitled “d synergistic process to enhance the bioavailability and bio efficacy of bioactive molecules", the entirety of which is expressly incorporated herein by referencePreamble to the Description

[0002] The following specification describes the invention and the manner in which it is to be performed:DESCRIPTION OF THE INVENTIONTechnical field of the invention

[0003] The present invention relates to a synergistic process to enhance the bioavailability and bio efficacy of bioactive molecules. More specifically, the process of the present invention integrates precision particle engineering of bioactive molecules along with the incorporation of synergistic molecules to enhance the solubility and hence bioavailability of the bioactive molecules.

[0004] Background of the invention

[0005] Bioavailability is the extent at which the drug enters the human circulation system and is available for action on the active site. Bioavailability is a parameter to judge the extent of a particular drug molecule’s absorption by the system and efficiency at the target site. Bioavailability is an integral part of theInternal Ref: OR26C001PCT01pharmacokinetics paradigm. The route of administration (ROA) and the drug dose can significantly impact both the rate and extent of bioavailability.

[0006] First pass metabolism refers to the metabolism processes a drug molecule undergoes as the bioactive compound passes through the liver and intestines before entering the bloodstream. First pass metabolism has a potential of breaking down the drug molecules, hence reducing their therapeutic effect. Many structural and physiological gastrointestinal (GI) alterations affect the absorption, typically by reducing bioavailability. Hence the enhancement of bioavailability of compounds is required while formulating new drugs.

[0007] Enhancement of bioavailability of the drug improves therapeutic effect at a smaller dosage making the drug more readily absorbed by the target site. By lowering the administered dosage, the side effects can also be reduced by a significant portion. There are multiple existing technologies for bioavailability enhancement of a drug.

[0008] Co-solvency is a method used to enhance bioavailability of adding a water miscible solvent to a poorly water soluble drug. Some examples of cosolvents are PEG 300, propylene glycol or ethanol. One of the major drawbacks of co-solvency is the toxicity due to the solvents used, especially when the drug is administered at high dosages. Transitioning the formulation to a solid oral dosage forms can be another drawback.

[0009] Complexation is a process by which cyclodextrins form inclusion complexes that increase their solubility. Cyclodexterins are capable of forming inclusion complexes by taking up a whole drug molecule into the cavity and affect the aqueous solubility and rate of dissolution. Cyclodextrins are expensive and have potential gastrointestinal side effects due to the high concentrations used for encapsulation.

[0010] Liposome formulations are another approach to enhancing the bioavailability of drug molecules. In this process, the drug molecules are incorporated in a phospholipid bilayer. They are widely used due to theirInternal Ref OR26C001PCT01biocompatibility with human systems and the potential to target specific tissues. One of the major drawbacks of liposomes includes chemical stability as phospholipids are susceptible to oxidations and hydrolysis that lead to premature degradation of the liposome structure. Liposomes can also trigger immune responses that might lead the biological system to reject the liposomes and trigger an allergic reaction.

[0011] Precision particle engineering is a branch of pharmaceutical development that involves obtaining optimal particle size and distribution. Modifying the size, shape, structure and surface properties of particles enhances their performance in drug formulations.

[0012] The precision particle engineering method increasing bioavailability tackles all the problems mentioned in the above existing technologies as no foreign substances are used for formulation of the drug molecules. This method also enhances solubility and absorption owing to the increased surface area. Size reduction is also a cost effective and economical method as compared to the existing technologies.

[0013] Molecular docking is the study of analysing the interaction between two or more molecular structures that allows us to identify target molecules. It allows locating the binding site of the molecules and assessment of the binding affinity of the molecule.

[0014] The patent application No. EP0012523A1 entitled “Therapeutic compositions with enhanced bioavailability and process for their preparation” discloses compositions of poorly soluble or water insoluble drugs and to compositions of relatively soluble drugs which have a tendency to agglomerate or crystallise in storage or after formulation into pharmaceutical dosage form. The new compositions provide higher dissolution rates of said drugs in vitro and increased bioavailability. The compositions of this invention comprise such poorly soluble, water insoluble or relatively soluble drugs, a non-toxic water soluble polymer and a wetting agent. A process for preparing the compositions is also disclosed. Compositions containing the known antifungal griseofulvin illustrate the invention.Internal Ref: OR26C001PCT01

[0015] The patent application No. IN202021044023A entitled “Preparation of universal bioenhancer blend” discloses compositions containing extracts and compounds obtained from the plant black pepper (Piper nigrum), long pepper (Piper longum) and ginger (Zingiber officinale) and Liquorice (Glycyrrhiza glabra), useful as bio-enhancers and bioavailability facilitators for a variety of molecules including anti-infective, anti-cancer agents and nutritional compositions. This invention also relates to method of preparation of universal bioenhancer blend which enhancing the biological availability, proficiency and effectivity of drugs, pharmaceuticals, nutraceuticals and other related compounds including amino acids, vitamins and other nutritional elements and ions.

[0016] Although various technologies exist in enhancing bioavailability, there exists certain limitations to them. The existing state of art is selective in nature and fails in addressing the concern of enhancing solubility, absorption and hence bioavailability. Thus, it is crucial to develop a process to enhance the bioavailability and bio efficacy of bioactive molecules.Summary of the invention

[0017] The present invention discloses a process for enhancing the bioavailability and bio-efficacy of bioactive molecules, particularly those exhibiting poor solubility, limited absorption, or rapid elimination from the body. The invention provides a systematic, reproducible, and scalable process that integrates computational screening, synergistic molecule selection, and precision particle engineering to improve the physicochemical and biological performance of bioactive molecules.

[0018] The process comprises initially extracting and isolating one or more bioactive molecules, followed by subjecting the isolated bioactive molecules to high-throughput screening using molecular modelling techniques. Based on the screening results, one or more synergistic auxiliary molecules capable of functionally enhancing the bioactive molecule are identified. The synergistic molecules are selected based on their ability to improve parameters such asInternal Ref: OR26C001PCT01solubility, stability, absorption, bioavailability, efficacy, and safety beyond additive effects.

[0019] The identified bioactive molecule and synergistic molecule are subsequently subjected to precision particle engineering, wherein physical characteristics including particle size, morphology, surface properties, and molecular association are controlled. This structured particle engineering facilitates solid dispersion, increases surface area, and enables improved solubility and absorption of the bioactive molecules. The process further includes mixing the bioactive and synergistic molecules to obtain a uniform mixture, followed by homogenization, preferably high-pressure homogenization, and spray drying to obtain a stable, free-flowing spray-dried powder.

[0020] The process of the present invention overcomes limitations associated with conventional bioactive formulations and enables targeted delivery, improved absorption, and enhanced bio-efficacy, making the resulting compositions suitable for use in nutraceutical, pharmaceutical, dietary supplement, and functional food applications.Brief description of the drawings

[0021] The foregoing and other features of embodiments will become more apparent from the following detailed description of embodiments when read in conjunction with the accompanying drawings. In the drawings, reference numerals refer to like elements.

[0022] FIG 1 illustrates a flowchart of a process to enhance the bioavailability and bio efficacy of bioactive molecules.

[0023] FIG 2 illustrates a flowchart of a process of preparation of quercetin complex to enhance bioavailability and bio efficacy.

[0024] FIG 3 illustrates a micrograph of quercetin complex after subjecting to scanning electron microscopy.Internal Ref: OR26C001PCT01

[0025] FIG 4 illustrates a micrograph of quercetin complex after subjecting to transmission electron microscopy.

[0026] FIG 5 illustrates a micrograph of quercetin complex after dynamic light scattering.Detailed description of the invention

[0027] In order to more clearly and concisely describe and point out the subject matter of the claimed invention, the following definitions are provided for specific terms, which are used in the following written description.

[0028] The term “Precision Particle Engineering” refers to the process of reducing a particle size to increase its surface area for better absorption and enhanced drug efficacy.

[0029] The term “Bioavailability” refers to the extent a substance or drug becomes completely available to its intended biological destination.

[0030] The term “Synergistic Effect” refers to the interaction or cooperation of two or more substances to produce a combined effect greater than the sum of their separate effects.

[0031] The term “Bio efficacy” refers to the effectiveness of the drug due to the bioavailability.

[0032] The present invention discloses a process to enhance the bioavailability and bio efficacy of bioactive molecules. The process comprises steps of isolating the bioactive molecules, high throughput screening of the biomolecules, identifying synergistic molecules, precision particle engineering of the bioactive molecules, homogenization of the mixture and spray drying the mixture. The process disclosed in the present invention facilitates incorporation of precision particle engineering to modify the physical characteristics of the bioactive molecule along with the addition of synergistic molecules to enhance the absorption. The process results in enhanced solubility, absorption hence increased bioavailability and efficacy of the bioactive molecules.Internal Ref: OR26C001PCT01

[0033] FIG 1 illustrates a flowchart of a process to enhance the bioavailability and bio efficacy of bioactive molecules. The process (100) starts at step (101) in which bioactive molecules are subjected to extraction and are hence isolated. At step (102), the isolated bioactive molecules are subjected to high throughput screening through molecular modelling. At step (103), relevant synergistic molecules are identified for the target bioactive molecule by the method of high throughput screening. The identification of appropriate synergistic molecules is a crucial factor in the process. One or more auxiliary molecules capable of exhibiting synergistic interaction with a selected primary bioactive molecule are identified using a computationally assisted high-throughput screening framework, integrating in silico, in vitro, and data-driven correlation models.

[0034] The screening framework involves systematic analysis of molecular descriptors, including but not limited to physicochemical properties, structural motifs, electronic parameters, conformational flexibility, and interaction propensity of a primary bioactive molecule and a plurality of candidate auxiliary molecules.

[0035] The computational chemistry techniques employed in this step include, but are not limited to molecular docking studies, molecular dynamics simulations, quantum chemical calculations, pharmacophore modeling, solubility and permeability prediction, and stability and degradation pathway analysis. These techniques are used to predict interaction potential, functional compatibility, and synergistic relevance between the bioactive molecule and auxiliary molecules.

[0036] Artificial intelligence (Al) and machine learning (ML) models are employed to correlate multi-parametric datasets derived from molecular interaction energies, predicted enhancement of bioavailability, stabilization effects, modulation of biological activity, reduction of irritancy or toxicity, and improvement in physicochemical performance. The Al and ML models are trained using a combination of historical experimental datasets, curated literature data, and internally generated screening data, enabling recognition of synergistic patterns that are not evident through conventional empirical screening methods.Internal Ref: OR26C001PCT01

[0037] According to the present invention, synergism is considered as a functional interaction wherein the auxiliary molecule enhances the efficacy, stability, delivery, bioavailability, safety, or performance of the primary bioactive molecule beyond a merely additive effect. The auxiliary molecules include but are not limited to bioavailability enhancers, stabilizing agents, penetration or absorption modulators, solubilizing agents, carrier-interactive molecules, metabolism-modifying agents, and surface-active or vesicle-forming compounds.

[0038] The output of this step comprises the selection of one or more synergistic auxiliary molecules, computationally validated to exhibit a high probability of functional synergy with the bioactive molecule.

[0039] At step (104), the bioactive molecule and the synergistic molecules identified are subjected to precision particle engineering where the physical characteristics including size, shape and surface properties are modified accordingly. Additionally, structured precision particle engineering facilitates solid dispersion aiding in increasing surface area of the bioactive molecules hence enhancing the solubility and bioavailability of the bioactive molecules. Structured engineering of bioactive molecules is further useful in combination therapy wherein combination therapy aids in reducing development of drug resistance.

[0040] Precision particle engineering techniques include, but are not limited to controlled precipitation processes, nano- or micro-structuring techniques, self-assembly-driven molecular association, controlled solvent evaporation, surfactant-mediated organization, and shear- or energy-assisted particle formation methods.

[0041] At step (105), bioactive molecules and the synergistic molecules are mixed to obtain a uniform mixture using synergistic formulation technique. The addition of synergistic molecules enhances the absorption of the bioactive molecule in turn enhancing the bioavailability and bio efficacy of the molecule. The application of precision particle engineering along with synergistic technology further aids in precise targeted delivery of the bioactive molecules.Internal Ref: OR26C001PCT01

[0042] At step (106), the mixture of bioactive molecules and synergistic molecules is subjected to homogenization. At step (107), the homogenized mixture comprising bioactive molecules and synergistic molecules is subjected to spray drying to obtain spray dried powder of the mixture.

[0043] The obtained mixture exhibits enhanced solubility hence exhibiting increased absorption and bioavailability. The process is compatible with bioactive active molecules including berberine, vitamin C, palmitoyl ethanol amide, resveratrol, quercetin, rutin, milk thistle etc. exhibiting a wide range of solubility.

[0044] The following examples are offered to illustrate various aspects of the invention. However, the examples are not intended to limit or define the scope of the invention in any manner.Example 1: A process for preparation of quercetin complex to enhance the bioavailability and efficacy of quercetin.

[0045] Quercetin complex is prepared using the process of the present invention to enhance the bioavailability and efficacy of quercetin. The present example iterates the process for extraction, purification, and complexation of quercetin with synergistic bioactive compounds, derived from Sophora japonica and citrus sources, to obtain a stable, enhanced-bioavailability quercetin complex.

[0046] The whole plant material of Sophora japonica is initially subjected to size reduction by crushing to obtain crushed raw material. The crushed material is then subjected to ethanolic extraction under controlled conditions to solubilize flavonoid constituents. The ethanolic extract thus obtained is subjected to acidification, followed by crystallization using ethanol to selectively precipitate quercetin. The crystallized product is subsequently dried to obtain purified quercetin.

[0047] In another embodiment, the flowers of Sophora japonica are subjected to alkaline extraction to obtain a miscella containing flavonoid glycosides. The miscella is filtered to remove insoluble residues, producing a crude extract.Internal Ref: OR26C001PCT01

[0048] The crude extract is then subjected to aqueous extraction at approximately 70°C, followed by concentration of the extract. The concentrated extract is spray-dried to yield a fine aqueous extract powder rich in synergistic phytoconstituents.

[0049] In a further embodiment, the filtered crude extract obtained from the alkaline extraction of Sophora japonica flower is subjected to controlled crystallization to isolate rutin. The rutin crystals are dried to obtain rutin powder of desired purity.

[0050] In another embodiment of the invention, dried orange peel is crushed and subjected to alkaline extraction. The extract is filtered to remove insoluble matter, yielding a crude extract. The crude extract is subjected to crystallization to isolate hesperidin, followed by drying to obtain hesperidin powder.

[0051] FIG 2 illustrates a flowchart of a process of preparation of quercetin complex to enhance bioavailability and bio efficacy. The process (200) of preparation of quercetin complex starts with step (201) by subjecting to milling the purified quercetin obtained as above in powder form from Sophora Japonica whole plant at a concentration of 80% w / w along with aqueous extract of Sophora Japonica at a concentration of 5% w / w and 5% w / w rutin bioactive isolated from Sophora Japonica flower to obtain a homogeneous blend.

[0052] At step (202), the resulting blend is mixed with synergistic ingredient, wherein the auxiliary bioactive enhance the functional performance of quercetin. The synergistic ingredient is identified using a high throughput screening,

[0053] At step (203), the mixture is subjected to complexation to form a synergistic molecular complex. At step (204), the complex thus formed is processed as a slurry and subjected to high-pressure homogenization at about 500 bar for three cycles.

[0054] At step (205), the homogenized mixture is subjected to molecular complexation with coconut-derived lipids. The coconut lipids comprise mediumchain triglycerides, phospholipids, or a combination thereof. The complexation is carried out by dispersing the synergistic quercetin composition into the lipid phase under controlled temperature and agitation conditions, resulting in the formation ofInternal Ref: OR26C001PCT01a uniform slurry wherein quercetin and associated bioactives are molecularly associated with the lipid matrix.

[0055] At step (206), the slurry obtained after lipid complexation is subjected to high-pressure homogenization at a pressure of approximately 500 bar, for three homogenization cycles. This step ensures uniform dispersion of the quercetin-lipid complex, reduction in particle size, formation of a stable molecular complex, and enhancement of solubility and bioavailability of quercetin.

[0056] At step (207), the homogenized slurry is then subjected to spray drying to convert the liquid dispersion into a dry, free-flowing powder. The spray drying is achieved by maintaining the inlet temperature at approximately 160°C -170°C, and the outlet temperature is maintained at approximately 90°C. The spray drying step encapsulates the quercetin-lipid molecular complex, resulting in improved stability, reduced hygroscopicity, and ease of handling.

[0057] The dried powder obtained after spray drying is collected and optionally sieved to obtain a uniform particle size distribution. The final product thus obtained is a quercetin complex from the process of the present invention is suitable for use in nutraceutical, pharmaceutical, or functional food formulations

[0058] Quercetin complex prepared using the process of the present invention is subjected to characterization by different instrum entational techniques using scanning electron microscopy (SEM) and Transmission electron microscopy (TEM) and evaluating Zeta potential and particle size distribution to confirm the structural morphology.Example 2: Characterization of quercetin complex using scanning electron microscopy.

[0059] Quercetin complex prepared using the process of the present invention is subjected to scanning electron microscopy. Quercetin complex is stored in doublesided carbon ribbon-wrapped aluminum stubs, sputter-coated with a thin layer of gold by a sputter gold coater and scanned at a 10 kV accelerating voltage.Internal Ref: OR26C001PCT01

[0060] FIG 3 illustrates a micrograph of quercetin complex after subjecting to scanning electron microscopy. The SEM micrographs reveal that the quercetin complex particles are predominantly nanosized and exhibit an approximately spherical morphology. The particles appear to be uniformly distributed, with minimal to no visible aggregation, indicating effective complexation and stabilization within the formulation. The observed morphology demonstrates the physical stability of the quercetin complex and support the suitability of the formulation for enhanced dispersion and bioavailability.Example 3: Characterization of quercetin complex using transmission electron microscopy

[0061] Quercetin complex prepared using the process of the present invention is subjected to transmission electron microscopy. Quercetin complex is dispersed in a suitable medium and subjected to sonication for approximately 10 minutes to obtain a uniform dispersion. A small quantity of the dispersed sample is then placed onto glow-discharged, thin carbon-coated TEM grids and allowed to dry at room temperature prior to imaging.

[0062] FIG 4 illustrates a micrograph of quercetin complex after subjecting to transmission electron microscopy. The TEM micrographs show that the quercetin complex is nanosized and exhibit a generally spherical morphology with a smooth surface. The particles are observed to be well dispersed, with no significant aggregation, indicating effective stabilization of the quercetin complex.

[0063] The TEM observations further support the structural uniformity and physical stability of the quercetin complex obtained by the process of the present invention.Example 4: Analysis of particle size and zeta potential of quercetin complex using dynamic light scattering.

[0064] Quercetin complex prepared using the process of the present invention is subjected to dynamic light scattering to analyse the particle size and zeta potential.Internal Ref OR26C001PCT01

[0065] The measurements are carried out using a dynamic light scattering in which quercetin complex is diluted with purified water to obtain suitable scattering intensity and are measured at a controlled temperature of approximately 25°C.

[0066] The particle size distribution is calculated based on the Stokes-Einstein equation, and the zeta potential is determined using Nano DTS software. Each measurement is performed using at least three independent sets, with six runs per set, to ensure reproducibility of the results.

[0067] Zeta potential is an indicator of the surface charge characteristics of dispersed particles and reflects the degree of electrostatic repulsion between particles, which in turn influences suspension stability. In general, zeta potential values greater than approximately ±30 mV are considered indicative of sufficient electrostatic stabilization to minimize aggregation.

[0068] FIG 5 illustrates a micrograph of quercetin complex after dynamic light scattering. Quercetin complex exhibited a negative surface charge, with a zeta potential value of approximately -31.68 mV, as shown in Figure 5 (a). This value indicates that the quercetin complex forms a physically stable colloidal system. The particle size distribution analysis revealed that the quercetin complex particles are within a size range of approximately 150 nm to 300 nm, with a mean particle size of about 214 nm, as illustrated in Figure 5 (b). The relatively narrow size distribution indicates a uniform dispersion of particles.

[0069] Mean particle size is an important parameter influencing the physical stability, solubility, release characteristics, and biological performance of nutraceutical and pharmaceutical formulations. The observed nanoscale particle size and uniform distribution of quercetin complex suggest that the formulation is suitable for enhanced delivery and absorption, while also indicating good formulation reproducibility and physical stability.

[0070] The results obtained from SEM, TEM, particle size distribution, and zeta potential analyses demonstrate that the quercetin complex prepared by the processInternal Ref: OR26C001PCT01of the present invention exhibits stable nanoscale characteristics and is suitable for incorporation into nutraceutical and functional food compositions.

[0071] The present invention discloses a process to enhance the bioavailability and bio efficacy of bioactive molecules. The process overcomes the limitations of reduced solubility and rapid elimination of bioactive molecules. The process discloses precision particle engineering of bioactive molecules along with the addition of synergistic molecules. The process aids in providing accurate targeted delivery of the bioactive molecule with specific shape, size and surface properties. The bioactive molecules obtained from the process disclosed pass through the blood brain barrier achieving targeted delivery. The process hence results in enhanced solubility and bioavailability of bioactives.

Claims

Internal Ref: OR26C001PCT01Claims:We Claim,1. A process to prepare a bioactive complex to enhance the bioavailability and efficacy of one or more bioactive molecules, the process (100) comprises the steps of:a. extracting one or more bioactive molecules from a natural source (ioi);b. subjecting the isolated bioactive molecules to high throughput screening through molecular modelling (102);c. identifying one or more synergistic molecules for the isolated bioactive molecule by the method of high throughput screening (103);d. subjecting the bioactive molecules and the identified synergistic molecules to precision particle engineering such that one or more physical characteristics such as size, shape and surface properties are modified (104);e. mixing the bioactive molecules and the synergistic molecules to obtain a uniform mixture using synergistic formulation technique (105);f. subj ecting the mixture of the bioactive molecules and the synergistic molecules to homogenization (106); andg. subjecting the homogenized mixture comprising bioactive molecules and synergistic molecules to spray drying to obtain spray dried bioactive complex powder.

2. The process as claimed in claim 1, wherein the bioactive molecule is selected from a group comprising a polyphenol, flavonoid, alkaloid, terpene, vitamin, mineral, peptide, or combinations thereof.

3. The process as claimed in claim 1, wherein the high-throughput screening using molecular modelling techniques comprises one or more of molecularInternal Ref: OR26C001PCT01docking, molecular dynamics simulations, quantum chemical calculations, pharmacophore modelling, solubility prediction, permeability prediction, and stability analysis.

4. The process as claimed in claim 1, wherein the synergistic auxiliary molecules comprise one or more of bioavailability enhancers, stabilizing agents, penetration enhancers, solubilizing agents, lipid-based carriers, metabolism-modifying agents, or surface-active compounds.

5. The process as claimed in claim 1, wherein the precision particle engineering step results in particles having a mean particle size in the range of 50 nm to 500 nm, preferably 150 nm to 300 nm.

6. The process as claimed in claim 1, wherein the precision particle engineering results in one or more particles having spherical or near- spherical morphology with controlled surface charge characteristics.

7. The process as claimed in claim 1, wherein homogenization is carried out using high-pressure homogenization at a pressure of 500 bar for three cycles.

8. The process as claimed in claim 1, wherein the spray-dried bioactive complex exhibits a zeta potential of -31.68 mV, indicating enhanced colloidal stability.

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