# US 2017/0281702 A1 · Method of isolating anti-viral ingredients from baphicacanthus cusia, compositions comprising them and their medical use

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- Number: US 2017/0281702 A1
- Document ID: US20170281702A1
- Status: Published application
- Kind: A1
- Application number: 15/089,687
- Filed: 2016-04-04
- Published: 2017-10-05
- Earliest claimed: 2016-04-04
- Assignees: Macau University of Science and Technology
- Applicants: Macau University of Science and Technology
- Inventors: Jing-Rong Wang; Zhi-Hong Jiang; Qi-Tong Feng; Guo-Yuan Zhu; Wei-Na Gao; Zifeng Yang; Nanshan Zhong
- Web page: https://patentgrep.com/patent/US20170281702A1

## CPC

- A61K 36/19: Acanthaceae (Acanthus family)
- A61K 31/404: Indoles, e.g. pindolol
- A61K 31/538: ortho- or peri-condensed with carbocyclic ring systems
- A61K 31/567: substituted in position 17 alpha, e.g. mestranol, norethandrolone
- A61K 2236/15: involving mechanical treatment, e.g. chopping up, cutting or grinding
- A61K 2236/17: involving drying, e.g. sun-drying or wilting
- A61K 2236/33: involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K 2236/55: Liquid-liquid separation; Phase separation
- 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)

## Abstract

The present invention provides a method of isolating at least one ingredient with anti-viral efficacy from Baphicacanthus cusia. The ingredient can be an alkaloid, a triterpenoid, a lignan, a phenylethanoid, a sesquiterpene lactone, or a flavonoid. Two new alkaloids are produced, which have not been previously reported. Moreover, the method isolates 12 compounds which could not or have not been previously isolated. A pharmaceutical composition includes the at least one ingredient and at least one pharmaceutical tolerable excipient. A method of treating a subject suffering from a viral disease includes administering at least one ingredient isolated from Baphicacanthus cusia.

## Claims

### Claim 1

A method of isolating at least one ingredient selected from the group consisting of an alkaloid, a triterpenoid, a lignan, a phenylethanoid, a sesquiterpene lactone and a flavonoid from Baphicacanthus cusia which method comprises steps of:
(i) subjecting Baphicacanthus cusia plant material to a solvent extraction with an extraction solvent for obtaining a crude extract, wherein the extraction solvent comprises an aliphatic alcohol and wherein the solvent extraction is carried out at temperatures above 40° C., and wherein the Baphicacanthus cusia plant material comprises roots;
(ii) contacting the crude extract with at least a first, a second and a third separation solvent for obtaining at least a first, a second and a third layer, wherein the first separation solvent comprises water, the second separation solvent comprises an ester and the third separation solvent comprises an aliphatic alcohol, and wherein the obtained first layer comprises the first separation solvent, the second layer comprises the second separation solvent and the third layer comprises the third separation solvent;
(iii) subjecting at least one of the second or the third layer to at least a first and a second chromatographic separation step.

### Claim 2 (depends on 1)

The method of claim 1, wherein the ingredient is selected from the group consisting of a benzoheterocyclic alkaloid, a quinoline alkaloid, an indole alkaloid, a triterpenoid, a lignan and a phenylethanoid.

### Claim 3 (depends on 1)

The method of claim 1, wherein the ingredient is selected from the group consisting of:
a compound of Formula (1)
a compound of Formula (2)
a compound of Formula (5)
a compound of Formula (6)
a compound of Formula (7)
a compound of Formula (8)
a compound of Formula (9)
a compound of Formula (11)
a compound of Formula (14)
a compound of Formula (19)
a compound of Formula (20)
a compound of Formula (21)
a compound of Formula (25)
and
a compound of Formula (26)

### Claim 4 (depends on 1)

The method of claim 1, wherein the Baphicacanthus cusia plant material essentially consists of roots of Baphicacanthus cusia, and wherein the method further comprises steps before step (i) of
a) drying, and
b) at least one of cutting, shredding, milling or pulverizing the roots.

### Claim 5 (depends on 1)

The method of claim 1, wherein the extraction solvent in step (i) comprises 75 vol. % to 85 vol. % aqueous ethanol and wherein the solvent extraction is carried out for at least 0.5 to 8 h at a temperature of at least 50° C.

### Claim 6 (depends on 1)

The method of claim 1, wherein the Baphicacanthus cusia plant material is refluxed with the extraction solvent and wherein the solvent extraction is carried out for three times and the amount of Baphicacanthus cusia plant material in relation to the total amount of extraction solvent is between 30 mg/ml and 50 mg/ml.

### Claim 7 (depends on 1)

The method of claim 1, wherein contacting the crude extract with the at least first, second and third separation solvent in step (ii) is carried out by
a) suspending the crude extract in the first separation solvent;
b) then adding the second separation solvent for forming the first and the second layer and separating the second layer; and
c) subsequently adding the third separation solvent to the first layer after step b) for forming a third layer,
and wherein the first separation solvent essentially consists of water, the second separation solvent comprises ethyl acetate and the third separation solvent comprises n-butanol.

### Claim 8 (depends on 1)

The method of claim 1, wherein the first chromatographic separation step in step (iii) is carried out with a classical column chromatography, and wherein the second separation step in step (iii) is independently selected from the group consisting of classical column chromatography, medium pressure liquid chromatography including preparative medium pressure liquid chromatography, high-performance liquid chromatography including preparative and semi-preparative high-performance liquid chromatography, and thin layer chromatography.

### Claim 9 (depends on 1)

A method of treating a subject suffering from a viral disease comprising administering an effective amount of at least one ingredient selected from the group consisting of an alkaloid, a triterpenoid, a lignan, a phenylethanoid, a sesquiterpene lactone and a flavonoid isolated from Baphicacanthus cusia according to the method of claim 1 to the subject.

### Claim 10 (depends on 9)

The method of claim 9, wherein the ingredient isolated from Baphicacanthus cusia is selected from the group consisting of:
a compound of Formula (5)
a compound of Formula (7)
a compound of Formula (9)
and
a compound of Formula (19)

### Claim 11 (depends on 9)

The method of claim 9 comprising administrating an effective amount of one ingredient isolated from Baphicacanthus cusia selected from:
a compound of Formula (7)
or
a compound of Formula (19)

### Claim 12 (depends on 9)

The method of claim 9, wherein the viral disease is caused by a RNA virus.

### Claim 13 (depends on 12)

The method of claim 12, wherein the RNA virus is Influenza A virus subtype H1N1.

### Claim 14

A method of treating a subject suffering from a viral disease comprising administering an effective amount of a compound having one of the following Formulas to the subject:
Formula (5)
Formula (7)
Formula (9)
or
Formula (19)

### Claim 15 (depends on 14)

The method of claim 14, wherein the compound has a Formula selected from:
Formula (7)
or
Formula (19)

### Claim 16 (depends on 14)

The method of claim 14, wherein the viral disease is caused by an Influenza A virus.

### Claim 17 (depends on 16)

The method of claim 16, wherein the Influenza A virus is selected from subtypes H3N2, H5N1 or H1N1.

### Claim 18 (depends on 14)

The method of claim 14, wherein the subject is a human and the amount of the compound administered is between 1 mg/kg and 50 mg/kg body weight.