(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)

(19) World Intellectual Property Organization

International Bureau
(10) International Publication Number
(43) International Publication Date Χ 1 /1 i . A t
29 December 2011 (29.12.2011) WO 2 l l /161 1 Al

(51) International Patent Classification: (74) Common Representative: VISHWANATH, Kannan;

BO 11/02 (2006.01) A61K 36/49 (2006.01)
(21) International Application Number:
PCT/IN20 11/000404
(22) International Filing Date:
16 June 201 1 (16.06.201 1)
(25) Filing Language: English
(26) Publication Language: English
(30) Priority Data:
1849/MUM/2010 22 June 2010 (22.06.2010)
(71) Applicant (for all designated States except US): AA-
JANEYA BIOTECH PVT. LIMITED [IN/—]; Aan-
janeya House, No. 34, Postal Colony, Chembur, Mumbai
400 071 (IN).
(72) Inventor; and
(71) Applicant : VISHWANATH, Kannan [IN/IN]; M/s.
Aaajaneya Biotech Pvt. Limited, Aanjaneya House, No.
34, Postal Colony, Chembur, Mumbai 400 071 (IN).
(72) Inventors: VISHWANANTHAN, Kashi; M/s. A aa
janeya Biotech Pvt. Limited, Aanjaneya House, No. 34,
Postal Colony, Chembur, Mumbai 400 071 (IN).
LATHA, C ; M/s. Aaajaneya Biotech Pvt. Limited, A an
janeya House, No. 34, Postal Colony, Chembur, Mumbai
400 071 (IN). THANGARAJ, Suresh; M/s. Aaajaneya
Biotech Pvt. Limited, Aanjaneya House, No. 34, Postal
Colony, Chembur, Mumbai 400 071 (IN).
M/s. Aaajaneya Biotech Pvt. Limited, Aanjaneya House,
No. 34, Postal Colony, Chembur, Mumbai 400 071 (IN).
(81) Designated States (unless otherwise indicated, for every
kind of national protection available): AE, AG, AL, AM,
AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ,
CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO,
DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT,
HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP,
KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD,
ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI,
IN NO, NZ, OM, PE, PG, PH, PL, PT, RO, RS, RU, SC, SD,
SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR,
TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW.
(84) Designated States (unless otherwise indicated, for every
kind of regional protection available): ARIPO (BW, GH,
GM, KE, LR, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG,
ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ,
TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK,
EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU,
LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK,
SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ,
GW, ML, MR, NE, SN, TD, TG).
Published:
— with international search report (Art. 21(3))
— before the expiration of the time limit for amending the
claims and to be republished in the event of receipt of
amendments (Rule 48.2(h))

- (54) Title: A DIRECT METHOD FOR PREPARING QUININE HYDROCHLORIDE FROM CINCHONA BARK
(57) Abstract: The disclosure of invention relates to a cost effective and pharmaceutically acceptable process for the direct extrac-
tion method for quinine hydrochloride from Cinchona bark. In the present invention, wherein a mixture of Cinchona alkaloids
were extracted with an organic solvent from Cinchona bark and the alkaloids were precipitated out from the acidified aqueous lay-
¾ er by adjusting optimal pH using NaOH solution. Precipitate was subjected to water purification and quinine hydrochloride was
crystallized at ambient temperature.
 P T/IN201 1/000404

Title: - A direct method for preparing quinine hydrochloride from Cinchona bark

Field of the Invention:-

Quinine (cinchonan-9-ol, 6'-methoxy-, (8. alpha. 9R)-) is an antiprotozoal and an

antimyotonic, and is known for the treatment of malaria caused by Plasmodium species,
the treatment and prophylaxis of nocturnal recumbency leg muscle cramps, and the
treatment of babesiosis caused by Babesia microti. Quinine is structurally similar to
quinidine, which is also an antiprotozoal, but can function as an antiarrhythmic. The said
Cinchona alkaloid isolated from the bark of several species of Cinchona trees, the
medicinally active bark, which is stripped from the tree, dried and powdered, contains a
variety of alkaloids having their antimalarial property. In 1820, quinine (( )-(6-
methoxyquinolin-4-yl) (( 2 S , 4S, 8R) - 8-vinylquinuclidin-2-yl) methanol) was identified
as an active compound. Since then, cinchona alkaloids especially, quinine have played a
pivotal medicinal role. Approximately 700 metric tons of cinchona alkaloids are now
extracted from the bark of Cinchona annually. Nearly half of this is used in the food and
beverages industry as a bitter additive and much of the remaining quinine and quinidine
is used as an important antimalarial drug and muscle relaxant compound and as a cardiac
depressant (antiarrhythmic).
Cinchona trees remain the only economically practical source of quinine.
However, under wartime pressure, research towards its synthetic production was
undertaken.

Prior art:-

A formal chemical synthesis was accomplished in 1944 by American chemists

R.B. Woodward and W.E. Doering. Since then, several more efficient quinine total
syntheses have been achieved, but none of them can compete in economic terms with
isolation of the alkaloid from natural sources.
U.S. Patent Application No.20090239900, which teaches more specifically, the
quinine or quinidine salt included in the combination product according to the present
invention may be any commercially available quinine or quinidine salt, or hydrate
thereof, such as but not limited to quinine sulphate, quinine sulphate dihydrate, quinine
monohydrochloride, quinine monohydrochloride dihydrate, quinine bisulfate, quinine
bisulfate heptahydrate, quinine hydrobromide, quinine dihydrochloride, quinine
dihydrobromide, quinidine hydrochloride, quinidine hydrochloride monohydrate and
quinidine sulfate dihydrate (cinquine).

CN 101402634 (A) the invention discloses a method for separating and purifying
alkaloid in Peruvian bark. By a method for separating and purifying quina
through soaking and extracting of acid aqueous solution and enriching of ion
exchange resin, the invention greatly reduces the application amount of organic
solvents, obviously reduces the energy consumption required by the
concentration of the organic solvents, greatly reduces the production cost on the
basis of ensuring that extraction yield rate is not lower than solvent method and
lightens the adverse effect on the environment. The method has the advantages
that the method changes extracting media, avoids the water extraction of the
organic solvents, adopts the repeatedly regenerated ion exchange resin as a
concentration and enrichment method, avoids the heating concentration and
reduces the energy consumption.

DE 3704850 (A1);- The invention relates to a process for the extraction of

quinine from ground cinchona by extraction with supercritical C02 at a pressure
of 100 to 500 bar and a temperature of 80 to 120@C. The extracted quinine is
precipitated as quinine sulphate in a separation vessel which is partly filled with
sulphuric acid.

GB 758173 (A), Quinine is extracted from the ground product obtained from the
alkaline maceration of cinchona bark by treating it with a solvent mixture of at
least one hydrocarbon and a chlorinated hydrocarbon, ketone or alcohol.
Preferred proportions are 5-22 parts by volume of the latter to 100 parts of the
hydrocarbon. Examples show the use of the following particular solvent mixtures:
85-95 per cent of naphtha with 5-15 per cent of isopropanol, 88-92 per cent of
turpentine with 8-12 per cent of isoamyl alcohol, 88-92 per cent of hydrocarbons
of B.Pt. 170-222 DEG C . (85 per cent aromatics) with 8-12 per cent of methyl
isobutyl carbinol, 82-88 per cent of xylene with 12-1 8 per cent of methyl ethyl
ketone, and 88-92 per cent of schist oil with 8-12 per cent of chloroform.
CN 101088999 (A) The present invention provides several processes of
preparing 3-amino quinine dihydrochloride. The first process includes the
oximation and reduction of 3-quininone hydrochloride material to obtain 3-amino
quinine, and the reaction with hydrochloric acid to form 3-amino quinine
dihydrochloride. The second process includes the direct reduction of 3-quininone
hydrochloride material in the presence of noble metal catalyst to obtain 3-amin
oquinine, and the reaction with hydrochloric acid to form 3-amino quinine
dihydrochloride. The third process includes the oximation, etherification and
reduction of 3-quininone hydrchloride material to obtain 3-amino quinine, and the
reaction with hydrochloric acid to form 3-amino quinine dihydrochloride. These
processes are simple, high in yield, low in production cost and suitable for use in
industrial production.

Objective of the present invention

An object of the present invention is that, there are no economically viable

methods for the preparation of quinine hydrochloride. The existing method for
manufacturing quinine hydrochloride is from the sulphate form of quinine. Quinine
hydrochloride via quinine sulphate increases the cost of production and loss of active
molecule.

Further very important object of the present invention is that the said invention

relates a method of preparing quinine hydrochloride directly from Cinchona bark is

commercially significant and great achievement which method is non - obvious in the
filed of alkaloid chemistry.

Summery of the invention,

Quinine is a basic amine and always presented as a salt. Various preparations of

quinine salt include the hydrochloride, dihydrochloride, sulfate, bisulfate and gluconate.
The most generally used form of quinine salt was the sulphate, but more recently the
hydrochloride has come into favor as it is much more soluble in water. The sulphate is
taken up only by 725 parts of water, while the hydrochloride is dissolved by 35 parts.
Each is readily dissolved with acid, bisulphate in 9 parts and the acid hydrochloride in 0.6
part of water. The presence of excess of acid is a disadvantage in intramuscular or
intravenous injection and especially the solubility of the neutral hydrochloride weighs
heavily in the favor of quinine hydrochloride.

Description of the invention

Quinine is a basic amine and always presented as a salt. Various preparations of

quinine salt include the hydrochloride, dihydrochloride, sulfate, bisulfate and gluconate.
The most generally used form of quinine salt was the sulphate, but alleged invention
relates to a direct method for preparation of quinine hydrochloride from cinchona bark
comprising the following steps, taking a known quantity of dried and pulverized
Cinchona bark is mixed with quicklime in the ration of 0.5: 0.9 preferably 1:0.4 and 6%
caustic solution was added to this mixture. The blend was thoroughly mixed and kept
aside for 6 to 12 hours preferably 8 to 10 hr. After the specified time the blend was
extracted with oreganice solvent selected from Toluene, chloroform, acetone, methanol,
ethanol, at the ratio of 0.5:6 preferably 1:5 at 40 to 60 degree temperature more
preferably at 50° C for 2 to 4 hours preferably for 3 hrs.

The second and third extraction was carried out with 0.5:4 more preferably 1:3
blend and Toluene ratio for 2 to 4 hours preferably 3 hrs separately. The extracts were
collected together and filtered through high flow bed. The filtrate was extracted with 5%
hydrochloric acid and the mixture was stirred properly for 10 to 40 minutes preferably
30 minutes. The solution was kept undisturbed until to get separate the toluene and
aqueous layer. When two layers were completely separated out, the aqueous layer was
collected and adjusted the pH 4 to 5 with using 5% NaOH. Maximum amount of
precipitate was allowed to form by adding NaOH solution. The crude precipitate of
various cinchona alkaloids was filtered and subjected to dry at 40° C using hot air.
 Crude precipitate and hot distilled water ( 1 :5) were slowly and continuously
stirred with 15% activated charcoal for 30 minutes at 65° C. The mixture was filtered hot
and the filtrate was continuously stirred in order to reach a temperature of 30° C.
Crystallization was observed at ambient temperature. The crystals were dried at 40° C
with hot air. When the moisture content was about 6 to 10% the crystals was subjected to
various analytical tests such as solubility, pH, SOR, and HPLC to conform national and
international pharmacopeias.

Example 1

Pulverized Cinchona bark of 100 gm was soaked with quicklime (2 gm) and 6-8%
KOH solution. The blend was vigorously mixed and kept undisturbed for 10-15 hr. The
above mixture was extracted continuously with Acetone of 800 ml, 200 ml, and 200 ml
respectively for 3 hr separately at 60° C . The extract was filtered, pooled together and
acidified with 100 ml of 2 % HCl and the mixture was stirred continuously to mix the
fractions properly. The solvent mixture was allowed to stand for 1 hr to get the layer
separated out. The aqueous layer was collected and the pH was adjusted to 4.5 to 5 with
3% LIQ NH3 solution to obtain precipitate. The precipitate was filtered and dried at a
temperature of 45 °C. Crude precipitate was added to five times quantity of distilled
water containing 20% activated carbon at 80° C. The mixture was continuously stirred for
30 minutes. The filtrate obtained at hot was continuously stirred for crystallization.
Crystallization was observed at 10-1 ° C temperature. The crystals were taken for various
analyses.

Example 2

Five hundred gm of Cinchona bark was mixed with milk of lime 150 m l and 3 to
5% caustic soda solution and the mixture was kept for 10 to 12 hr after mixing properly.
The mixture was extracted with MeOH (1.5 1) for 9 hr at 70° C. After filtering the extract,
the extract was acidified with 1% HCl and the two layers were allowed to separate it out.
The aqueous layer pH was adjusted to 4.5 to 5.5 with 1% NaOH solution to obtain crude
precipitate. After the filtration the precipitate was dried at 45°C. Crude precipitate was
dissolved in distilled water ( 1 :5). Activated charcoal was added to the mixture at 65° C.
The mixture was gradually and continuously stirred for 30 minutes and filtered hot.
Crystals were obtained at ambient temperature. The crystals were subjected for various
analyses.
What is Claimed,

. A direct method for preparation of quinine hydrochloride from cinchona

bark comprising the following steps,

i. Taking a known quantity of dried and pulverized Cinchona

bark, mixed with quicklime in the ratio of 0.5: 0.9 and 6%
caustic solution was added to this mixture.
ii. Blend obtained at step (i) was thoroughly mixed and kept
aside for 6 to 12 hours after the specified time the blend was
extracted using organic solvents at the ratio of 0.5:6 at 40 to 60
degree temperature for 2 to 4 hours preferably ,
iii. The second and third extraction was carried out using 0.5:4
blend and solvent ratio for 2 to 4 hours separately; the extracts
were collected together and filtered through high flow bed.
iv. The filtrate obtained at step (iii) was extracted with 5%
hydrochloric acid and the mixture was stirred properly for 10 to
40 minutes, the solution was kept undisturbed until to get
separate organic and aqueous layer, when two layers were
completely separated out,
v. The aqueous layer obtained at step iv was collected and
adjusted the pH 4 to 5 using 5% NaOH. Until to get maximum
amount of precipitate was formed by addition of NaOH
solution,
vi. The crude precipitate obtained at above step of various
cinchona alkaloids was filtered and dried at 40° C using hot air,
vii. To above resultant crude precipitate added hot distilled water
( 1 :5) slowly under constant stirring with 15% activated
charcoal for 30 minutes at 65° C,
vii. The reaction mixture obtained at step vii allowed to filtered hot
and the filtrate was continuously stirred until to reach a
 temperature of 30 C, followed by crystallization at ambient
temperature, and dried at 40° C with hot air,

2. A direct method as claimed in claim 1, wherein the organic solvents

used are selected from toluene, methanol, ethanol, chloroform,
acetone,more particularly toluene,

3. A direct method as claimed in claim 1, wherein the Cinchona bar is

mixed with quicklime in the ration of 0.5: 0.9 preferably 1:0.4,

4. A direct method as claimed in claim 1, wherein the blend was thoroughly

mixed and kept aside for 6 to 12 hours preferably 8 to 10 hr,

5. A direct method as claimed in above claims, wherein the blend was

extracted with Toluene at the ratio of 0.5:6 preferably 1:5 at 40 to 60
degree temperature more preferably at 50° C for 2 to 4 hours preferably
for 3 hrs,

6. A direct method as claimed in claim 1, wherein the second and third

extraction was carried out using 0.5:4 more preferably 1:3 blend and
Toulene ratio for 2 to 4 hours preferably 3 hrs separately, followed by
Alteration at high flew bed,

7. A direct method as claimed in claim 1, wherein the filtrate was extracted

with 5% hydrochloric acid and the mixture was stirred properly for 0 to 40

minutes preferably 30 minutes,

8. A direct method as claimed in claim 1, wherein the two layers were

completely separated out, and the aqueous layer was collected and
adjusted the pH 4 to 5 using 5% NaOH, precipitate was formed by adding
NaOH
 solution, said crude precipitate cinchona alkaloids was filtered and dried
using hot air, at 40° C ,

9 . A direct method as claimed in above claims , wherein the crude precipitate

and hot distilled water ( 1 :5) were added slowly under constant stirring using
15% activated charcoal for 30 minutes at 65° C,

10. A direct method as claimed in claim 1 and 9 wherein the s filtered hot and the

filtrate was continuously stirred until to reach a temperature of 30° C the

crystals were dried at 40° C with hot air,
 International application No

PCT/IN2011/0O0404
A . CLASSIFICATION O F SUBJECT MATTER
INV. B01D11/02 A61K36/49
ADD.

According to International Patent Classification (IPC) or to both national classification and IPC

B. FIELDS SEARCHED
Minimum documentation searched (classification system followed by classification symbols
B01D A61K

Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched

Electronic data base consulted during the international search (name of data base and, where practical, search terms used)

EPO-Internal

C . DOCUMENTS CONSIDERED TO BE RELEVANT

Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.

GB 758 173 A (BUZAS ANDRE) 1-10

3 October 1956 (1956-10-03)
the whole document

□ Further documents are listed in the continuation of Box C . See patent family annex.

* Special categories of cited documents :

"A" document defining the general state of the art which is not
considered to be of particular relevance
"E" earlier document but published on or after the international
filing date
"L" document which may throw doubts on priority claim(s) or
which is cited to establish the publication date of another
citation or other special reason (as specified)
"O" document referring to a n oral disclosure, use, exhibition or
other means
"P" document published prior to the international filing date but
later than the priority date claimed
"T" later document published after the international filing date
or priority date and not in conflict with the application but
cited to understand the principle or theory underlying the
invention
"X" document of particular relevance; the claimed invention
cannot be considered novel or cannot be considered to
involve an inventive step when the document is taken alone
" document of particular relevance; the claimed invention
cannot be considered to involve an inventive step when the
document is combined with one or more other such docu¬
ments, such combination being obvious to a person skilled
in the art.
"&" document member of the same patent family

Date of the actual completion of the international search Date of mailing of the international search report

16 November 2011 22/11/2011

Name and mailing address of the ISA/ Authorized officer
European Patent Office, P.B. 5818 Patentlaan 2
NL - 2280 HV Rijswijk
Tel. (+31-70) 340-2040,
Fax: (+31-70) 340-3016 Haderlei n , Andreas
 International application No
Information on patent family members
PCT/IN2011/0OQ404
Patent document Publication Patent family Publication
cited in search report date member(s) date

GB 758173 03-10-1956 NONE