Mixed micelles for drug delivery

Abstract

The present invention provides very small sized (even as low as average particle size of 10- 50nm) and highly permeable mixed micelles for enhanced and controlled drug delivery including solubility and bioavailability.

Description

[0001] M5069-00001

[0002] MIXED MICELLES FOR DRUG DELIVERY FIELD OF THE INVENTION:

[0003] The present invention provides very small sized (even as low as average particle size of 10-50nm) and highly permeable mixed micelles for enhanced drug delivery including solubility and bioavailability.

[0004] Current invention is versatile with respect to nature of active (small molecules, peptide and protein drugs, phytochemicals, natural extracts both of hydrophilic or hydrophobic nature) to be incorporated; amount of active (up to 20% w / v); and its nanosize.

[0005] BACKGROUND OF THE INVENTION:

[0006] The following background discussion includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.

[0007] Presently, various advanced drug delivery systems such as micelles, nanogels, nanocrystals, nanotubes, and nanocapsules are employed for the delivery of drugs and other therapeutically active molecules. These nano-scale delivery systems are increasingly preferred over conventional dosage forms as they address several inherent limitations associated with traditional drugs, including poor aqueous solubility, low biostability, dose-related toxicity, inadequate bioavailability, and multidrug resistance. Among these systems, micellar carriers are particularly well recognized for their ability to solubilize poorly water-soluble drugs through the formation of hydrophobic cores, thereby facilitating efficient drug loading and targeted delivery. This targeted transport minimizes drug loss and systemic side effects while enhancing drug availability at the desired site of action.

[0008] Polymeric micelles, in particular, are considered effective drug delivery vehicles owing to their nanoscale size, structural stability, surface characteristics, and enhanced permeability and retention (EPR) effect. These micelles are formed through the self-assembly of amphiphilic block copolymers into a characteristic core-shell architecture, wherein the hydrophobic core serves as a reservoir for lipophilic drugs and the hydrophilic shell stabilizes the system in aqueous environments. Typically, polymeric micelles possess sizes ranging from 10 to 100 nm, which is ideal for improved permeation and retention at target tissues.

[0009] At low concentrations, amphiphilic surfactant or polymer molecules remain dispersed in solution as monomers. Upon reaching a threshold concentration, known as the critical micelle concentration (CMC), these molecules spontaneously self-aggregate to form micellar structures. The CMC is a critical parameter influencing the stability, drug loading capacity, and performance of micellar systems.

[0010] Mixed micelles represent an advancement over conventional polymeric micelles, wherein at least one of the amphiphilic components is lipidic in nature. The incorporation of lipidic cosurfactants results in enhanced thermodynamic stability and improved versatility of the micellar system, enabling efficient encapsulation of both hydrophilic and lipophilic drugs. Such systems exhibit higher drug loading capacity, improved encapsulation efficiency, and superior delivery performance compared to single-component micellar formulations.

[0011] Despite these advantages, currently known micellar and mixed micellar formulations suffer from several drawbacks. For instance, DE102018219633A1 discloses an aqueous cosmetic cleaning composition comprising specific surfactants and lipophilic substances, wherein theM5069-00001

[0012] surfactant concentration is equal to or greater than that of the lipophilic substances. However, the high surfactant content in this formulation poses a significant risk of skin irritation, thereby limiting its suitability for therapeutic topical applications.

[0013] EP316411 IB 1 describes an aqueous composition comprising poloxamer-based micelles encapsulating hydrophobic agents. While effective for solubilizing hydrophobic drugs, the formulation involves complex preparation steps, requires high concentrations of poloxamers, and is limited to loading only hydrophobic actives. Furthermore, the process necessitates strict temperature control, making it inefficient and less suitable for scalable manufacturing.

[0014] Similarly, KR20230106361 A discloses mixed micellar systems that require elevated temperatures and multi-step processing methods. Such conditions increase energy consumption and operational complexity, rendering the process unsuitable for large-scale or cost-effective production.

[0015] US5654337A describes pharmaceutical formulations containing lecithin and surfactants at substantially high concentrations, with lecithin levels reaching up to 30% and surfactant concentrations up to 20%. These formulations do not claim or demonstrate nano-scale micellar systems and employ relatively low water content (25-60%), relying heavily on non-aqueous solvents such as ethyl alcohol, benzyl alcohol, and isopropyl myristate. High excipient load and solvent dependency limit formulation flexibility, patient acceptability, and safety.

[0016] EP2211837B1 discloses a non-aqueous solution of diclofenac diethylamine intended for transdermal delivery using high quantities of volatile organic solvents. Upon application, rapid solvent evaporation leads to skin dryness, reduced permeation, and compromised skin hydration. To counteract these effects, additional penetration enhancers, humectants, and urea are required, increasing formulation complexity and cost. Repeated application of such solventrich systems can disrupt the stratum corneum, weaken the skin barrier, and increase susceptibility to infections and dermatological disorders.

[0017] Accordingly, there exists an unmet need for a stable, low- surfactant, energy-efficient, and scalable mixed micellar drug delivery system capable of encapsulating both hydrophilic and lipophilic actives, providing enhanced permeation and controlled release, while minimizing skin irritation, formulation complexity, and manufacturing constraints.

[0018] OBJECTIVE OF THE INVENTION:

[0019] The primary object of the present invention is to overcome the drawbacks associated with prior art.

[0020] Another object of the present invention is to provide a composition with enhanced drug delivery, permeability and bioavailability for both hydrophilic and hydrophobic drugs and BCS class I to IV drugs.

[0021] Another object of the present invention is to provide a composition for encapsulating various drugs.

[0022] Another object of the present invention is to provide a stable composition with smaller particle sizes.

[0023] Another object of the present invention is to provide a composition with high drug loading capacities, high Encapsulation Efficiency.

[0024] Another object of the present invention is to provide a composition with uniform particle morphology for consistent delivery and stability.

[0025] Another object of the present invention is to provide a composition with robust lipid / surfactant-based formulation for modem medical applications.M5069-00001

[0026] Summary of the Invention:

[0027] The Invention provides a mixed micellar pharmaceutical or cosmeceutical composition for delivery of therapeutic agents, comprising:

[0028] (a) a lipidic co-surfactant selected from phosphatidylcholine or lecithin of natural or synthetic origin comprising Lipoid S 75, S 100, 90H, 90G, 100H and / or Phosal MCT 53 (53% phosphatidylcholine in medium-chain triglycerides (MCT)) in an amount of 0.5-20% w / w;

[0029] (b) a surfactant selected from polysorbates, sorbitan esters, Cremophor RH 40 (hydrogenated castor oil PEG 40) or combinations thereof present in an amount of 0.1- 25% w / w;

[0030] (c) one or more organic solvents selected from ethanol, isopropyl alcohol, methanol, benzyl alcohol, butanol, acetonitrile, ethyl acetate or mixtures thereof in an amount of 2-25% w / w;

[0031] (d) at least one active pharmaceutical ingredient selected from analgesic, antiinflammatory, neuropathic pain-relieving, antimicrobial, anticancer, cardiovascular, ocular, anti-VEGF, vitamin, nutraceutical, cosmetic or peptide-based agents or pharmaceutically acceptable salts thereof in an amount of 0.1-20% w / w; and

[0032] (e) water q.s. to 100% w / w, wherein the components spontaneously self-assemble to form nano-sized mixed micelles having an average particle size of less than 100 nm. In an embodiment, the mixed micelles have an average particle size of 20-50 nm, a poly dispersity index of not more than 0.35, a zeta potential between ±0.5 mV and ±25 mV, a drug entrapment efficiency of at least 80%, and a drug content of at least 90%.

[0033] In an embodiment, the organic solvent content is reduced or eliminated when the active pharmaceutical ingredient is hydrophilic, enabling formation of solvent-free mixed micelles. In an embodiment, the composition comprises one or more optional therapeutic or sensory agents selected from methyl salicylate, menthol, thymol, linseed oil, lavender oil, mentha oil, castor oil, sunflower oil, mustard oil or camphor, without adversely affecting micelle formation or stability.

[0034] In an embodiment, the mixed micelles are incorporated into a dosage form selected from a gel, hydrogel, cream, lotion, spray, inhalation, douche, enema, foam or solution.

[0035] In an embodiment, the composition is prepared by a low-energy and scalable process comprising dissolving the lipidic co- surfactant, surfactant, organic solvent and active pharmaceutical ingredient in an organic phase, introducing the organic phase into an aqueous phase under controlled stirring or microfluidic mixing, and allowing spontaneous selfassembly of mixed micelles at temperatures not exceeding 40°C.

[0036] In an embodiment, the composition is used for the manufacture of a medicament for management of pain, inflammation, arthritis, neuropathic pain or other inflammatory conditions.

[0037] DETAILED DESCRIPTION OF DRAWINGS:

[0038] To further clarify the advantages and features of the present invention, a more particular description of the invention will be rendered by reference to specific embodiments thereof, which are illustrated in the appended drawings. It is appreciated that these drawings depict only typical embodiments of the invention and are therefore not to be considered limiting in their scope. The invention will be described and explained with additional specificity and detail with the accompanying drawings in which:

[0039] Figure 1: (a) scanning electron microscopy (SEM) and (b) transmission electron microscopy (TEM) image of developed MMs illustrating relatively uniform surface of well -dispersed spherical to irregular micelles.M5069-00001

[0040] Figure 2 Cumulative in vitro drug release (a and c) (IVRT) and permeation (2b and d) (IVPT) across Wistar rat skin, profiles of various formulations of diclofenac diethyl amine (DDEA; 2a 6 b) and Amitriptyline (AMT; 2c & d) with time. Fig 2(e) shows confocal microscopy images describing in vivo permeation of FITC-loaded micelles through shaved dorsal skin surface of rats at 30 min, 2 h, and 8 h post application to live animals in comparison to free FITC.

[0041] Marketed (blue): Volitra APS manufactured by Troika Pharmaceuticals Ltd. and marketed by Sun Pharma Industries Ltd; Formulation (orange): DDEA loaded mixed micelles; Suspension (gray): free DDEA suspended in 1% CMC All data are presented as mean ± SD (n=6).

[0042] Figure 3 Analgesic (a and b) and anti-inflammatory (c and d) activity of developed DDEA 4% nanospray and marketed Volitra APS treatment versus no treatment, in terms of: (a) acute thermal analgesia describing spinal, reflex mediated pain response at 30 minutes post application, using the tail flick test; and (b) complex supraspinal thermal analgesia at 30 minutes post application, using the hot plate test; and (c) croton oil-induced right ear oedema followed by respective treatment 30 minutes post induction and observation in terms of increase in weight of treated ear; and (d) its histopathology at 8 h following treatment represented as H&E-stained ear tissue sections at 400X. A- Positive control, B- Marketed, C-Formulation, D-Naive groups

[0043] Arrows and circles- Red- epidermis, Blue- Inflammatory infiltrate, Green- Dermis.

[0044] 50pl dose of 4% DDEA loaded nanomicelles was applied per rat in each experiment.

[0045] Figure 4 Anti-arthritic activity of micellar formulations of DDEA (a-d), and AMT (e-h) in the Complete Freund’s Adjuvant (CFA)-induced (day -7) inflammatory arthritis in rats and its comparison with other treatments. Treatments were started at day 0 and effects recorded at day 7 and 14 following treatment and expressed as: (a, g) % recovery in inflammation and oedema measured as paw volume using a Plethysmometer; (b, h) total white blood cell (WBC) count at day 14, describing inflammation (reduction in WBC levels compared to the disease control indicates attenuation of the inflammatory response); (c, f) supraspinal thermal analgesia using hot plate method; and (d) mechanical analgesia using the Randall-Selitto paw pressure test. Positive: CFA-induced arthritis but no treatment; oral diclofenac: peroral administration of an equivalent amount of powdered diclofenac tablet (Reactin® 50 mg Marketed by Cipla pharma and life sciences ltd) corresponding to 5.14 mg / Kg body weight; Dynapar®: 50 pl of 4.64% DDEA (equivalent to 4% diclofenac sodium) spray manufactured and marketed by Troika Pharmaceuticals Ltd., Ahmedabad, India; Omnigel®: lOOmg gel containing 1.16% DDEA + 10% methyl salicylate + Menthol 5% + linseed oil 3% (manufactured by Metrocraft, Baddi and marketed by Cipla Health Ltd., India ); 1% and 2% gel: Both are gel formulations (lOOmg each) containing 1.16 and 2.32% DDEA loaded MMs, respectively and 10% methyl salicylate + 5% Menthol in each case; 2% spray: 50 pl of MM dispersion loaded with 2.32 % DDEA+ 10% methyl salicylate + Menthol 5%; 4% spray: 50 pl of MM dispersion loaded with 4.64% DDEA; AMT-MMs cream: lOOmg of cream incorporating AMT (5% w / w) loaded MMs; AMT oral: peroral administration of an equivalent amount of powdered AMT tablet (Tryptomer®-lOmg Dr Reddy’s laboratory limited) corresponding to 61.7 mg / Kg bodyweight.

[0046] Figure 5 (a, c) Representative hind paw images of rats showing anti -arthritic activity of various DDEA (a) and AMT(c) treatments in the CFA-induced arthritis model as explained above in fig 4. Progressive swelling, erythema, and joint deformity were observed in CFA control animals. Captured Images illustrate changes in paw swelling and inflammation across various treatments, (b, d) Representative radiological (X-ray) images illustrating joint integrity and bone architecture in various groups, highlighting the protective effect of micellar formulations against arthritis-induced joint damage.

[0047] Figure 6 In vivo investigation describing modulation of key pro-inflammatory and neuropathic pain markers in serum of rats, including TNF-a (inflammation signalling), IL-7 (acute-phase response), IL- ip (pyrogenic), substance P (pain transmission) and CRP (systemicM5069-00001

[0048] inflammation) following topical treatment with various formulations of DDEA (a-e) and AMT (f-j) in the CFA-induced arthritis model described previously in Fig 4.

[0049] The Dunnett' s T3 multiple comparisons test results indicate significant differences between all treatment groups and the control group. Most of the comparisons show strong statistical significance (p < 0.0001), with a few exceptions such as "Disease control vs. Omnigel" and "Oral Diclofenac vs. 2% gel," where no significant differences were found. The adjusted p-values further confirm the robustness of the observed differences.

[0050] Similarly, AMT MMs cream significantly reduced all markers compared to the disease control group, with p-values < 0.0001, as confirmed by Tukey's post hoc analysis. AMT MMs cream demonstrated superior efficacy over oral AMT, showing significant reductions in TNF-a (p = 0.0026), IL-6 (p < 0.0001), IL-1 (p < 0.0001), Substance P (p < 0.0001), and CRP (p < 0.0001). These findings suggest that AMT MMs cream effectively modulates both inflammatory responses and neuropathic pain, highlighting the potential of this nanoformulation as a localised treatment for neuropathic pain, with minimal or no side effects as associated with oral AMT administration.

[0051] Figure 7 shows Capsaicin-induced neuropathic pain animal model. The representative hind paw images of rats are shown in (a) and their response to thermal pain (b, supraspinal hot plate; c, acute and spinal tail flick) and cold allodynia (d) in capsaicin-induced neuropathic pain model in Wistar rats.

[0052] Timelines: -0.25 h (naive baseline), 0 h (capsaicin injection), 0.25 h (treatments initiated). Topical 5% w / w AMT-MMs cream demonstrated significantly higher analgesic efficacy compared to the disease control and oral AMT groups (p < 0.05), providing rapid and sustained reversal of induced neuropathy, demonstrating its potential as a localised and targeted delivery.

[0053] DETAILED DESCRIPTION:

[0054] For the purpose of promoting an understanding of the principles of the invention, reference will now be made to the embodiment illustrated in the drawings 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 normally occur to one skilled in the art to which the invention relates.

[0055] 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.

[0056] The present invention provides a mixed micelles formulation that significantly enhances the delivery and bioavailability of both hydrophilic and hydrophobic drugs. The formulation consists of one self-emulsifying / co-surfactant lipid compound (Lipoid S 75, S100, 90H, 90G, phosal MCT 53 / 53% phosphatidylcholine in medium-chain triglycerides (MCT)) coupled with stabilizers / solubilisers / emulsifiers (hydrogenated castor oil PEG 40, Gelucire 44 / 14, low and high molecular weight PEG, pol oxamer 188, Vitamin E, vitamin E derivatives) and an alcohol solvent. It features particles under 100 nm with near neutral zeta potential, achieving 50-100% Encapsulation Efficiency (EE). In vitro drug release is extended up to 72 hours. The drug release for mixed micelles was 90-100%. TEM and FESEM analyses reveal uniformly spherical to irregular particles. This versatile formulation supports high drug loading capacities (10-250% relative to lipid content) and offers a promising platform for improving pharmaceutical treatments.M5069-00001

[0057] In an embodiment, the invention provides a mixed micelle formulation for enhancing the delivery and bioavailability of hydrophilic and hydrophobic drugs, comprising an emulsifier blend combined with stabilizers and solubilisers for enhanced formulation.

[0058] The composition comprises following components:

[0059] a) At least a self-emulsifying lipid compound especially phosphatidyl choline or lecithin (of egg, soyabean or any other suitable origin) selected from the group comprising of Lipoid S 75, S100, 90H, 90G,100H, phosal MCT 53 (53% phosphatidylcholine in medium-chain triglycerides (MCT);

[0060] b) stabilizers / solubilisers / emulsifiers (hydrogenated castor oil PEG 40, Gelucire 44 / 14, low and high molecular weight PEG, polyols, poloxamer 188, Vitamin E, vitamin E derivatives) and

[0061] c) an aqueous / alcohol(s)-water / alcohol(s) / organic solvent.

[0062] The composition provides various technical advantages like:

[0063] • Utilizes a unique emulsifier blend combined with stabilizers and solubiliser / s for enhanced formulation.

[0064] • Exhibits prolonged drug release, with around 85% cumulative release over 72 hours • Features uniform spherical / irregular particles of 10-100 nm average size, contributing to consistent drug delivery and formulation stability.

[0065] The composition has various advantages as mentioned below over other known alternatives including imported alternatives:

[0066] • Effectively delivers both hydrophilic and hydrophobic drugs.

[0067] • Provides enhanced stability with particles under 100 nm and near neutral zeta potential.

[0068] • Achieves high drug content and encapsulation efficiency (75-100%).

[0069] • Offers superior prolonged and controlled release up to > 3 days

[0070] • Ensures uniform spherical to irregular particles for consistent delivery.

[0071] • Supports high drug loading capacities up to 30-50% with respect to the matrix that includes surfactant and lipid co surfactant.

[0072] • Utilizes effective solubility enhancers for better encapsulation and permeation.

[0073] • Maintains stability and mostly a near neutral zeta potential, preventing aggregation.

[0074] • Features advanced formulation technology for improved drug delivery.

[0075] • Enhanced skin absorption when applied topically to achieve quick relief

[0076] • Encapsulation in micelles reduces skin irritation of any irritating actives

[0077] • Can be used for routes including topical, transdermal, vaginal, rectal, parenteral, oral, ocular, otic, nasal, buccal or oral cavity, inhalation, nose to brain delivery

[0078] MMs loaded with suitable analgesics / anti-inflammatory actives offer:

[0079] • Targeted Pain Relief: Addresses local and systemic pain and inflammation

[0080] • Relieve neuropathic pain

[0081] • Safe and Effective Topical Application: An alternative to oral NSAIDs that show significant side effects

[0082] Various experiments were done for the selection of emulsifiers / stabilizers and solubilisers:

[0083] A series of surfactants, lipidic cosurfactants and solubilisers viz Cremophor RH 40, Polyethylene glycol 400, polyethylene glycol 600, polyethylene glycol 4000, polyethylene glycol 6000, vegetable-derived medium-chain triglycerides(MCTs) and fatty acid esters viz.M5069-00001

[0084] Captex 1000 (Glyceryl Tricaprate) and Captex 200 (Propylene Glycol Esters), Phosal 53 MCT (53% phosphatidylcholine in medium-chain triglycerides (MCT), Ethanol, Methanol, polyols, benzyl alcohol, isopropyl alcohol, and Acetonitrile were selected for the formulation of the current invention.

[0085] The concentration used for all the selected surfactants was above the critical micellar concentration. Different responses were taken into consideration like sedimentation and turbidity and particle size, stability, and drug loading for selection of the best formulation for further studies.

[0086] Procedure for preparing mixed micelle formulations

[0087] 1. Preparation and Measurement of Ingredients

[0088] 1.1. Calculate the required quantities of each ingredient based on the percentage composition specified in the formulation table and the desired final batch size.

[0089] 1.2. Weigh or measure all ingredients accurately.

[0090] 2. Preparation of MMs by preparing and mixing two phases

[0091] 2.1. Phase One (Organic Phase):

[0092] - Includes all ingredients that are soluble in organic or green solvents (e.g., ethanol, methanol, benzyl alcohol, acetonitrile, or other specified organic solvents, polyethers, polyols).

[0093] Stir (100-2000 rpm) and heat gently, if necessary, to a temperature in the range of 0- 50°C (as required by the solubility or volatility constraints of the ingredients).

[0094] 2.2. Phase Two (Aqueous Phase):

[0095] - Includes all ingredients that are soluble in water (e.g., water-soluble APIs, buffers, or excipients). Stir (100-2000rpm) and heat gently to 0-50°C (or to a temperature suitable for dissolving the water-soluble components).

[0096] - Note: If the active pharmaceutical ingredient (API) is water-soluble, it may be added directly to the aqueous phase without using organic solvent phase.

[0097] 2.3. While continually stirring, add the organic phase all at once into the aqueous phase. The rapid addition helps to form mixed micelles or nanoemulsion-type dispersions. Continue stirring for at least 5 minutes (or longer if required) to ensure a uniform dispersion and complete mixing.

[0098] 3. Cooling and Packaging

[0099] 3.1. Allow the final formulation to cool to room temperature (if heated).

[0100] 3.2. Transfer the final product into appropriate, airtight, and light-protective containers (as necessary based on stability requirements).

[0101] 3. Storage

[0102] - Store the final formulation in a cool, dry place, protected from light, and at conditions suitable for maintaining product stability (e.g., ambient or refrigerated depending on the API / excipient requirements).

[0103] Mixed Micelles produced using a scalable process:

[0104] Microfluidic Device Setup:

[0105] • A microfluidic device with a T-junction (or Y-junction, X-junction, Co-flow junction, Flow-focusing junction, H-junction, multi-inlet star junction, Star junction, treeM5069-00001

[0106] junction) was set up to facilitate efficient mixing of immiscible phases (ethanol and water).

[0107] • The organic phase (containing Cremophor RH40, Phosal MCT 53, and diclofenac diethylamine) was introduced into one inlet channel of the microfluidic device.

[0108] • The aqueous phase (water) was introduced into the other inlet channel.

[0109] Microfluidic Mixing:

[0110] • Syringe pumps were used to control the flow rates of both phases. The flow rate for the organic phase was set at 0.1-0.5 mL / min, and the aqueous phase at 1-2 mL / min to generate the required shear forces for micelle formation.

[0111] The two phases met at the T-junction, where rapid mixing occurred, resulting in the formation of mixed micelles as the fluids passed through the device

[0112] Formation of Mixed Micelles:

[0113] • The mixture self-assembled into mixed micelles under the shear forces generated in the microfluidic device.

[0114] • The mixed micelles were collected in a receiving reservoir after exiting the microfluidic device.

[0115] Post-mixing Hydration:

[0116] • After exiting the microfluidic device, the mixture was allowed to hydrate for 1 hour at room temperature to ensure complete micelle formation and equilibration.

[0117] Optimization:

[0118] • The flow rates, surfactant-to-phospholipid ratio, and diclofenac diethylamine concentration were optimized based on the desired micelle properties, such as drug release rate, micelle size, and stability.

[0119] • The process was scaled up using larger microfluidic devices or operating multiple devices in parallel if needed.

[0120] GENERAL FORMULATION TABLE 1:

[0121] One or more of the following ingredients and actives may be used to prepare Mixed Micelles, however one lipidic co-surfactant and one amphiphilic surfactant is always included to prepare MMs

[0122]

[0123] M5069-00001

[0124]

[0125] GENERAL FORMULATION TABLE 2:

[0126]

[0127] Formula 1

[0128]

[0129] Formula 2

[0130]

[0131] M5069-00001

[0132]

[0133] Formula 3

[0134]

[0135] Formula 4 (Nose to Brain Delivery)

[0136]

[0137] FORMULA-5

[0138]

[0139] Formula 6 (Ocular)

[0140]

[0141] Note on Notation

[0142] • “QS” (quantum satis) means “as much as needed to achieve the desired final quantity or volume.”

[0143] Incorporation of MMs into suitable Gel / hydrogel system as a secondary vehicle for ease of application where required:

[0144] Suitable gelling agent(s) (1-10 %) is mixed with an equal quantity of MMs incorporating 2x active to result in the final desired concentration

[0145] • The mixture is slowly stirred using a mechanical stirrer at a low speed for 15-30 minutes to ensure uniform distribution and prevent air entrapment.

[0146] Post-Preparation and Storage:

[0147] • The final gel formulation is allowed to sit overnight at room temperature for complete mixing and gelation.

[0148] • The prepared gel was stored in an airtight container to avoid evaporation and contamination.M5069-00001

[0149] The Greenness of the process

[0150] The developed mixed micellar formulation process demonstrates a high degree of energy efficiency and sustainability due to the absence of intensive thermal or high-shear processing steps. The process is conducted at mild temperatures not exceeding 40 °C and relies solely on controlled stirring (100-2000 rpm), thereby significantly reducing energy consumption compared to conventional high-energy homogenization or ultrasonication methods for preparation of nanoparticles. The use of low-shear mixing conditions enables uniform micelle formation without compromising physicochemical stability or drug encapsulation efficiency. Furthermore, the elimination of heating, high pressure, or multiple processing steps minimizes operational complexity and material degradation. Overall, the process offers a scalable, cost- effective, and environmentally benign approach for mixed micelle production with minimal energy input.

[0151] Example: Diclofenac diethylamine (DDEA) loaded mixed micelles (MMs) Characterization :

[0152]

[0153] FESEM and HR-TEM images of MMs indicated uniformly dispersed spherical / irregular particles with smooth surface. The micelles exist singly (no aggregates), show uniform size and even distribution

[0154] In-Vitro Release Test (IVRT) and In Vitro Permeation Test (IVPT) studies as per USFDA DDEA loaded micelles showed controlled release of 98.18±0.68% at 72 hours, while the market product (Volitra APS by Troika Pharmaceuticals Ltd. marketed by Sun Pharma Industries Ltd) released only 38.78±2.27% diclofenac in the same time

[0155] Kinetic Analysis showed the MMs followed the Korsmeyer-Peppas model confirming diffusion and polymer matrix relaxation-controlled release.

[0156] IVPT studies

[0157] The total drug permeated from DDEA loaded micelles is 830 and 1720% more than that of the marketed and free suspension.

[0158] Proof of Concept Study describing in vivo permeability of Fluorescent probe (FITC) labelled MMs when applied to the shaved dorsal skin surface of rats

[0159]

[0160] M5069-00001

[0161] Study describes dynamic penetration of micelles into the skin, confirming potential for rapid onset (within 30 minutes) and long-lasting effect of >8h

[0162] Activity in Rats

[0163] Analgesic activity was 250% to 270% better than that of the marketed comparator product.

[0164] Anti-Inflammatory Activity in Croton oil ear oedema model: DDEA micelles showed complete inhibition (98.27%) of inflammation versus only 55.32% inhibition of inflammation with the marketed (Dynapar®, manufactured and marketed by Troika Pharmaceuticals Ltd) comparator product. Notable decrease or complete absence of tissue inflammation and damage was observed in the treated ear tissue in histopathology studies (H&E, 400X) as described below:

[0165] • Positive (inflammation induced; no treatment) control- Tissues showed hyperkeratosis and acanthosis in epidermis with normally arranged collagen bundles in the pan dermis with dense inflammatory infiltrate and skin adnexal structures.

[0166] • Marketed (Dynapar® treated post inflammation) - Examined ear skin tissue showed normal thickness of epidermis with normally arranged collagen bundles in the pan dermis and moderate inflammatory infiltrate, mild oedema and skin adnexal structures (sebaceous glands and hair follicles).

[0167] • Formulation (nanomicelles treatment after inflammation induction) - skin tissue exhibited normal thickness of the epidermis and normally arranged collagen bundles in the pan dermis. No inflammation seen

[0168] • Naive- Section examined showed skin tissue having a normal thickness of epidermis with normally arranged collagen bundles in the pan dermis. No inflammation seen Antiarthritic Activity in Complete Freund's Adjuvant (CFA) induced joint inflammation in rats- Adult Wistar rats (200-230 g) were acclimatised with free access to food and water. Experimental protocols were approved by the IAEC, Panjab University, Chandigarh (PU / US / 99 / CPCSEA / IEAC / 2025 / 1023) and chronic inflammatory arthritis was induced by a single intraplantar injection of Complete Freund’s Adjuvant (CFA; 0.5 mg Mycobacterium butyricum in 0.1 mL paraffin oil) into the left hind paw 7 days prior to start of treatment which was continued for 14 days twice / day for DDEA mixed micelles and once a day for AMT mixed micelles.

[0169] Results were evaluated in terms of:

[0170] Paw volume / oedema (Plethysmometer) (p<0.00001)

[0171] 4% DDEA micellar spray showed significantly better recovery of 78.83% while marketed 4% DDEA spray Dynapar® (manufactured and marketed by Troikaa pharmaceuticals Ltd. Gujrat India) showed 5 times lesser response of just 15.35%.

[0172] WBC Count (p<0.00001)

[0173] WBC levels were completely recovered with the 2% and 4% spray formulations restoring values close to the naive group. Overall, the results demonstrate effective suppression of inflammation by the test formulations.

[0174] Supraspinal Thermal analgesia (Hot Plate) (p<0.00001) The spray formulations produced the highest pain relief outperforming oral and gel formulations in reducing thermal nociception. Effect was >2 times more pronounced than market formulation (Dynapar®).

[0175] Mechanical allodynia (Randall Selitto Test) (p<0.00001)

[0176] Both spray (4% and 2%) formulations showed highest recovery (>3 times than marketed product).

[0177] Radiology Reports (fig. 5b):M5069-00001

[0178] • Naive- No sign of arthritis and inflammation in joints

[0179] • Positive- Severe Inflammation in knee joints and metatarsal showing severe arthritis and low bone density

[0180] • Oral diclofenac - Mild arthritis in knee joints

[0181] • Marketed- Severe arthritis and inflammation seen in knee joints

[0182] • Omnigel® - Severe degradation changes in knee joints and metatarsal with low bone density

[0183] • 1% Gel -Slight arthritis in knee joint and slight loss of bone density

[0184] • 2% Gel - Mild arthritis in metatarsal and mild inflammation in knee joints

[0185] • 4% Nanospray- No arthritis only mild inflammation is seen

[0186] • 2% Nanospray- Mild Arthritis and very mild inflammation is seen in knee joints Thus, it can be concluded that both 4% and 2% diclofenac nanospray effectively reduced arthritis and joint inflammation, with 4% showing superior protection. They outperform oral diclofenac and the marketed product; ELISA studies also confirm their superior antiinflammatory effects

[0187] Pharmacokinetic Activity:

[0188] Single topical dose plasma pharmacokinetic studies were conducted in rats and promising results were obtained with significant longer ti / 2 and higher bioavailability indicating transdermal activity. Similar is also concluded from Pharmacodynamic studies where significant downregulation of inflammatory markers was observed in plasma.

[0189] Example: Amitriptyline (AMT) loaded MMs incorporated in a cream base (AMT MMs cream )

[0190] o Development and characterisation of AMT loaded MMs:

[0191]

[0192] o In vitro drug release test (IVRT):

[0193] The IVRT study (fig.2(c)) confirmed a marked time- and formulation-dependent profile for evaluated systems over 72 h. Unencapsulated AMT in cream base displayed a pronounced burst effect, releasing nearly 80-100% of the drug within 24 h, indicative of limited diffusional control. In contrast, AMT MMs and AMT -MMs cream both showed extended release, with 60-70% drug being released at 24 h and 95-100% by 72 h. Kinetic analysis showed that AMT MMs followed the Korsmeyer-Peppas model, consistent with diffusion-dominated, anomalous transport, while AMT-MMs cream obeyed second-order kinetics, reflecting a concentration-dependent, sustained release. The unencapsulated AMT cream was best described by first-order-like kinetics, explaining its rapid, gradient-driven burst release which will show poor capacity to maintain therapeutic levels over time.

[0194] o In vitro permeation test (IVPT):M5069-00001

[0195] The IVPT results show (fig.2(d)) distinct permeation profiles with AMT MMs (blue) exhibiting the fastest and most efficient drug permeation, with approximately 60-70% of the applied drug being permeated within 70 hours, ensuring quick therapeutic action. In contrast, the AMT MMs cream (orange), achieved only 25-30% permeation at 70 hours, probably due to the second diffusion barrier created by the cream base, making it ideal for prolonged therapeutic effects. The Unencapsulated AMT in Cream base (grey) showed lowest flux with only 10-15% of the drug permeation by the same time, reflecting a less efficient permeation profile.

[0196] Overall, the MMs exhibited fastest flux indicating that more AMT enters systemic circulation and is cleared from the body; while the AMT MMs cream formulations will provide a more sustained local effect and slower transdermal absorption, ensuring long-term therapeutic effects.

[0197] o In Vivo evaluation of AMT-MMs cream for analgesic and anti-inflammatory activity • Mechanical pain: CFA lowered paw-pressure thresholds to 113 g by day 14. AMT- MMs cream restored thresholds by 17.6% (day 7) and 62.6% (day 14), exceeding oral diclofenac and oral AMT.

[0198] • Thermal pain: Hot-plate latency increased 23.5 % at day 7 and 64.7 % at day 14 with AMT-MMs, the best among all other treatments.

[0199] • Paw volume: CFA increased paw volume to 5.47 mL; AMT-MMs reduced it by 33.5 % by day 14, close to the response shown by oral diclofenac (40.1 %).

[0200] • Leukocyte count: Elevated WBCs (14.8 x 103 / pL) were lowered to 9.7 x 103 / pL (~86 % recovery) with AMT-MMs, like oral diclofenac, indicating significant lowering of inflammation.

[0201] Topical 5 % AMT-MMs cream exhibited potent analgesic and anti-inflammatory effects, outperforming oral amitriptyline and approaching the efficacy of oral diclofenac in CFA-induced arthritic rats.

[0202] Topical 5 % w / w AMT-MMs cream also provided rapid and sustained relief of neuropathic pain, effectively reversing capsaicin-induced thermal hyperalgesia, spinal reflex pain, and cold allodynia. Its efficacy was comparable to or greater than oral AMT, highlighting the potential of this nano-formulation as a localised treatment for neuropathic pain, with minimal or no side effects significantly associated with oral AMT administration.

Claims

M5069-00001We Claim:

1. A mixed micellar pharmaceutical, nutraceutical or cosmeceutical composition for delivery of actives, comprising:(a) a lipidic co-surfactant selected from phosphatidylcholine or lecithin of natural or synthetic origin comprising Lipoid S 75, S 100, 90H, 90G, 100H and / or Phosal MCT 53 (53% phosphatidylcholine in medium-chain triglycerides (MCT)) present in an amount of 0.5-20% w / w;(b) a surfactant selected from polysorbates, sorbitan esters, Cremophor RH 40 (PEG- 40 hydrogenated castor oil) or combinations thereof present in an amount of 0.1-15% w / w;(c) one or more organic solvents selected from ethanol, isopropyl alcohol, methanol, benzyl alcohol, butanol, acetonitrile, ethyl acetate or mixtures thereof in an amount of 2-25% w / w;(d) at least one active selected from analgesic, anti-inflammatory, neuropathic pain- relieving, antimicrobial, anticancer, cardiovascular, flavors, perfumes, vitamins, phytochemicals, natural extracts, cosmetic or peptide-based agents or pharmaceutically acceptable salts thereof in an amount of 0.1-10% w / w; and(e) water q.s. to 100% w / w, wherein the components spontaneously self-assemble to form nano-sized mixed micelles having an average particle size of less than 100 nm.

2. The composition according to claim 1, wherein the mixed micelles have an average particle size of 20-50 nm, a poly dispersity index of not more than 0.35, a zeta potential between ±0.5 mV and ±25 mV, a drug entrapment efficiency of at least 70%, and a drug content of at least 90%.

3. The composition according to claim 1, wherein the organic solvent content is reduced or eliminated when the active pharmaceutical ingredient is hydrophilic, enabling formation of solvent-free mixed micelles.

4. The composition according to any one of claims 1, further comprising one or more optional therapeutic or sensory agents selected from methyl salicylate, menthol, mentha oil, linseed oil, thymol, camphor, eucalyptol, Peppermint oil, Spearmint oil, Capsicum oleoresin, Lemon oil without adversely affecting micelle formation or stability.

5. The composition according to claim 1, wherein the mixed micelles are incorporated into a dosage form selected from a gel, hydrogel, cream, lotion, spray, foam, inhalation or solution.

6. The composition according to claim 1, wherein the composition provides controlled and sustained drug release for 24-120 hours with enhanced permeation and reduced local irritation.

7. The composition according to claim 1, prepared by a low-energy and scalable green process comprising dissolving the lipidic co- surfactant, surfactant, organic solvent and active pharmaceutical ingredient in an organic phase, introducing the organic phase into an aqueous phase under controlled stirring or microfluidic mixing, and allowing spontaneous self-assembly of mixed micelles at temperatures not exceeding 40°C.

8. The composition according to claim 7, wherein the mixed micelles are prepared using a microfluidic device comprising a T-junction or Y-junction, X-junction, co-flow junction, Flow-focusing junction, H-junction, Multi-inlet star junction, Star junction, Tree junction with controlled organic-to-aqueous phase flow rates.

9. Use of the composition according to any one of claims 1-8 for the manufacture of a medicament for management of pain, inflammation, arthritis, neuropathic pain or other inflammatory conditions.M5069-0000110. Use of the composition according to any one of claims 1-8 for the manufacture of a medicament for topical, transdermal, vaginal, rectal, parenteral, oral, ocular, otic, nasal, nose to brain, buccal or oral cavity, oropharyngeal, nasopharyngeal and inhalation administration.

Images