# US 10,945,969 B2 · Compositions containing thymohydroquinone and their method of preparation

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- Number: US 10,945,969 B2
- Document ID: US10945969B2
- Status: Granted
- Kind: B2
- Application number: 16/232,197
- Filed: 2018-12-26
- Published: 2019-06-27
- Granted: 2021-03-16
- Earliest claimed: 2017-12-27
- Assignees: Sami Labs Limited
- Applicants: Muhammed Majeed; Kalyanam Nagabhushanam; Beena Bhat
- Inventors: Muhammed Majeed; Kalyanam Nagabhushanam; Beena Bhat
- Web page: https://patentgrep.com/patent/US10945969B2

## CPC

- A61K 31/05: Phenols {(cannabinoids A61K31/658)}
- A61K 31/065: Diphenyl-substituted acyclic alcohols
- A61K 31/122: having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
- A61K 31/19: Carboxylic acids, e.g. valproic acid (salicylic acid A61K31/60)
- A61K 31/191: having two or more hydroxy groups, e.g. gluconic acid
- A61K 31/704: attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin {(digitoxin A61K31/7048)}
- A61K 36/71: Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
- A61K 47/12: Carboxylic acids; Salts or anhydrides thereof
- A61K 47/14: Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
- A61K 47/22: Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 39/06: Free radical scavengers or antioxidants
- B01D 11/0203: {with a supercritical fluid}
- B01D 11/0288: {Applications, solvents}
- A61K 2236/00: Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- B01D 11/0211: {in combination with an electric or magnetic field}
- B01D 11/0292: {Treatment of the solvent}
- Y02P 20/54: using solvents, e.g. supercritical solvents or ionic liquids
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28)
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

Disclosed are compositions enriched with thymohydroquinone, further comprising of thymoquinone, hederagenin and/or α-hederin formulated by blending the active molecules isolated from the seeds of Nigella sativa. Also disclosed are novel processes for the isolation of bioactive components thymohydroquinone, thymoquinone from the seeds of Nigella sativa. A process for the isolation α-hederin and hederagenin from the spent material of Nigella sativa is also disclosed herein.

## Claims

### Claim 1

A composition comprising thymohydroquinone as represented by STR#1 isolated from the seeds of Nigella sativa, wherein said composition is standardised to contain 2%-5% w/w thymoquinone, 0.01%-10% w/w thymohydroquinone, 20%-95% w/w fatty acids, 0.001%-3% w/w α-hederin, 0.1%-4.0% w/w rosmarinic acid and 0.2%-2% w/w piperine.

### Claim 2 (depends on 1)

The composition as in claim 1, wherein the composition is prepared using a process comprising steps of:
o) Powdering the seeds of Nigella sativa, pulverizing using 1.5 mm mesh and passing through magnetic separator to obtain a coarse powder;
p) Subjecting the powder of step a) to super critical fluid extraction (SCFE) using liquid CO2 to obtain three fractions: low volatile compounds—S1 fraction, high volatile compounds—S2 fraction and spent residue;
q) Identifying the compounds in low volatile and high volatile fractions as thymoquinone as represented by STR#2, thymohydroquinone as represented by STR#1 and free fatty acids using Gas Chromatography with total yield of about 10-40% in S1 fraction and about 1-6% in S2 fraction respectively;
r) Extracting the spent residue of step b) with 5 volumes of ethanol at 60°−65° C. with stirring for 3 hours;
s) Filtering the concentrating the ethanol extract of step d) to obtain a brownish paste;
t) Dissolving the brownish paste of step e) in water and partitioning thrice with 3 volumes of hexane to obtain aqueous and hexane fractions;
u) Partitioning the aqueous fraction of step f) thrice with n-butanol;
v) Quenching the n-butanol fraction of step g) with water to strip off the solvent while maintaining 20-30% of total dissolved solids;
w) Spray drying to obtain pale brownish powder identified as α-hederin (CAS no. 27013-91-8) represented by STR#3 by HPLC with yield of 0.001-5% on a dry basis;
x) Blending the S2 fraction of step c) with α-hederin of step i) to obtain a mixture comprising thymohydroquinone, thymoquinone, α-hederin and free fatty acids;
y) Dissolving the mixture of step j) with ethanol in a reactor with stirring at 65°-70° C. for 30 minutes to obtain a homogenous mixture;
z) Stripping off the ethanol from mixture of step k) under vacuum and filtering to remove insolubles to obtain a brownish yellow oily liquid;
aa) Adding stabilizing agents (rosmarinic acid) and bioavailability enhancers—pieprine to the mixture of step l)
bb) Formulating the mixture of step m) into tablets, capsules, soft gels, powder, pills, syrups, lozenges, suspension, emulsions.

### Claim 3 (depends on 1)

The composition as in claim 1, wherein the free fatty acids in the compositions comprises of less than 0.5% w/w Ω 3 (omega 3) fatty acids, 40%-70% w/w Ω 6 (omega 6) fatty acids and 15%-25% w/w Ω 9 (omega 9) fatty acids.