AU4183078A ANTI-INFLAMMATORY AGENTS

Data originating from sources other than the EPO may not be accurate, complete, or up to date.

pr nz ii,1 QkV -FORMI Regulation 9 COMMONWEALTH OF AUSTRALIA PATENTS ACT, 1952-1973

APPLICATION FOR A PATENT 41830 /78 X/We, W. RAYMOND WALKER and BARRY BOETTCHER of 12
Mahogany Drive, New Lainbton, New South Wales, Australia hereby apply for the grant of a Patent for an
invention cntitled Anti- inflammatory Agents" which is described in the accompanying provisional
specification.

idy/Our address for service is Messrs GRIFFITH, HASSEL FRAZER, Patent Attorneys, of 323 Castlereagh
Street, S3YDNEY, New South Wales 2000, Commonwealth of Australia.

DATED this 25th day of September r 19 81 1K RAYMOND WALKER and ~CGST~ $~DBARRY BqETTCHER by
orPe of IFFITH, HASSEL FRAZER Fellows. Institute of Patent Attorneys of Australia.

To: The commissioner of Patents Commonwealth of Australia SEC SEE FOLIQOA..C N&W.E UJK .CiW _rU
4. 6 RE4 A. LFAI COMMONWALTH OF' AUSTRALIA PATENTS ACT 1952 (AS AMENDED) DECLARATION IN
SUPPORT OF AN APPLICATION FOR A PATENT (Name of applicant) (Title) (Futll name of signatory)
(Address of signatory) i(InserL detaii-s of invarnto/s) (insert details of assignment, etc.) paragraphs 3 an'i
4 for NonConvz,-tion applicatio.i) In support of an Application made by: ALCUSAL INCORPORATED
PTY.LTD. 41830 /78 for a patent for an invention entitled: "Complexes of Bivalent Copper" I, BARRY
BOETTCHER of 12 Mahogany Drive, New Lambton, New South Wales, Australia, do solemnly and
sincerely declare as follows:⟨br/⟩ 1. 1 am authorised by the above mentioned applicant for the patent to
make this Declaration on its behalf.

2. The name and address of each actual inventor of the invention is as follows:⟨br/⟩ Barry Boettcher, of
12 Mahogany Drive, New Lambton, New South Wales, Australia; William Raymond Walker, of
Department of Chemistry, University of Newcastle, Newcastle, New Southi Wales, Australia;⟨br/⟩ and
Michael Wel~e~ley Whitehouse, of Department of Experimental Pathology, Australian National
University, Canberra, A.C.T., Australia and the fact(x) upon~ which the applicant is entitled to make this
application are as folows:⟨br/⟩ Applicant is the assignee of the said invention from the said inventors by
Assignment Deed dated 27 August 1981 2w-- .Th .fPa-~a 5 .s .y .Ben .of .he ws 4 Ti& -bfte i- -a -re-f ced-
the -Preee!&i-- rpa," ejmph -fti-s44e~w~-(wefe)---the s~ ~~ii s-md-~av~t~-i reepect- th- 411er*ie -the-
tdb-je{ h-epv+e (Place and date of signing) Declared at Sydney this /40 day of P 19 81.

Signed:⟨br/⟩ Position: (A GRIFFITH HASSEL FRAZER, BOX 2133, G.P.O. SYDNEY, NSW 2001 AUSTRALIA (12)
AUSTRALIAN PATENT ABRIDGMENT (19) AU (11) AU-B] -41 830/78 (54) (71) (21) (23) (43) (51 )2 (72
COMPLEXES OF BIVALENT COPPER ALCUSAL INCORPORATED PTY. LTD.

41 830/78 520 726 (22) 28.11.77 22. 11 .78 (24) 28.11.78 28.6.79 (44) 25.2.82 C07C 65/U4 C07D 239/48
C07C 239/42 BARRY BOETTGHER, WILLIAM RAYMVOND WALKER AND MICHAEL WELLLSLEY
WHITEHOUSE 74) LUH (57 tUsE of the compounds as anti -i nfl ammatory agents is also cl aime~d.

Cl aim Neutral copper (I j) comipl exes characterized in that they have the formul a Cu[C 6H 4 X)COOl 2
ROH wherein ROH represents an al kanol and wherein X can be located in any posi tion and is SH, SnH or
NHR 1 where R1 i s N\ H2 cool, N1 L'OOl! COOH ./2 D92 *142/1E (11) AU-B1-41 830/78 2 rfNH COOH N/
COOH NN COOH N D1,1 COOH COOH NYCl CH 3 *142/19 U. ASTAL IA ⟨2 Form PATENTS ACT 1952-1973
COMPLETE SPECIFICATION (ORIGINAL) Class:⟨br/⟩ Int. Cl:⟨br/⟩ FOR OFFICE USE 4183078*swctafs 49.

Application Number:⟨br/⟩ Lodged:⟨br/⟩ Complete Specification-Lodged:⟨br/⟩ Accepted:⟨br/⟩ Published:

⟨br/⟩ PRiority:⟨br/⟩ (I Related Art:⟨br/⟩ Namne of Applicant Address of Applica Act~jalIlnventor:⟨br/⟩
Addre;3s for Servic TO BE_-COMPLETED BY APPLICANT 2~*L Y C A t BARR DOTTCIE~pWILLAM ~AY1ODW-
1 ""JER~qLLZEY WIIIIUE 12, Mahogany Drive, New Lambton, New Sout'l Wales, 2305;⟨br/⟩ nt: Det--f-Ge
ISty Newca244-= tie V tUe, N6 ~hW~e 3~8 and Dapt...o Exeim a 26G0.

-jpo 4 I1 t e:⟨br/⟩ BARRY BOETTCHER; WILLIAM RAYMOND WALKER;⟨br/⟩ MICHAEL WELLESLEY

WHITEHOUSE i fllww V 1 y A Griffith, H-assel Frazer, 323 Castlereagh St., SYDNEY N.S.W. 2000 AUSTRALIA
Complete Specification for the invention entitled: "COMPLEXES OF BIVALENT COPPER" loigstatement is a
full description' of this invention, including the best method of performing it known -1This invention
relates to novel anti-inflammatory copper complexes and to anti-inflammatory compositions and
processes utilizing such complexes.

Many studies of the role of copper, and especially complexes of copper, in the treatment of inflammatory
diseases have been reported in recent years. One of the g most recent studies describes the use of
parenterally 0 administered copper complexes, and particularly copper (II) 00 salicylate, in the treatment
of rheumatic disease (Sorenson e1 and Hangarter, Inifammnation, 2, 217-2'8 (1977)).

The prior art describes a wide variety of coppersalicylate complexes prepared by the reaction of
inorganic copper (II) salts with salicylic acid in aqueous solution.

The specific complex formed in aqueous solution has been found to depend on the pH of the aqueous
solution and both neutral complexes such as the pale blue crystalline bis (salicylato) copper (lI)
tetrahydrate OH 0 0-Cu -0 H 2 0 2 n i*ic^wn and anionic complexes such as the olive green sodium
salicylatocuprate (II) 0 c o v-11u0 jj 0 are known in the art.

It is an object of this invention to provide novel neutral copper complexes prepared by the reaction
of/copper (i.r) compounds and substituted benzoic acids in non-aqueous solvents. A further object of
the invention is to provide a method for the treatment of inflammatory diseases of animals by the
administration of compositions comprising as active ingredient the novel copper complexes of the
invention.

Accordingly in one/embodiment the invention provides neutral copper complexes of the formula Cu[C
6H4 (X)COO]2 ROH wherein ROH represents an alkanol, preferably a C1 to C6 aliphatic alkanol, and
wherein X can be located in any position and is OH, SH, SeH, NH 2 or NHR where R is:⟨br/⟩ 3 I*rsP~L"Y I I
"Rq OR I/ COOH COOH N OR NH COOH COOH O7R N COOHOR I CH3 13CH 3 The composition also
comprises a pharmaceutically acceptable non-aqueous carrier.

Certain of the complexes of the above formula are novel compounds and form a further aspect of the
present invention.

Although in no way wishing to be bound by theory, as a result of X-ray studies the complexes of the
invention have been shown to have the following dimeric structure:⟨br/⟩ 4 -1 ~*YY~ur~ _I-YIY Li_ LLILI ~L
ROH x 0 C u 0 0 C 0 0 HOR (1 4a -he/ preferred compound arcording to the iv n is the copper salicylate
complex which' i-sr believed -t-have the formula:⟨br/⟩ ROH OH 0 I _/I O-Cu C, HO 1 0 00 O HOR The
compounds of the invention may be prepared by reacting inorganic copper (II) compounds, for example
copper salts or cupric hydroxide, with the appropriate substituted benzoic acid in the presence of an
alkanol. For example, reaction of cupric hydroxide with salicylic acid in anhydrous ethanol gives the
complex of formula Cu[C6 H (OH)COO] 2 C2H5OH, hereinafter referred to by the formula Cu[H Sal] 2
C2H5OH, which forms deep green crystals when crystallised from anhydrous ethanol containing excess
(approx. 5M) salicylic acid.

The compounds of the invention have been found to be particularly useful in the treatment of
inflammatory diseases in animals. Thus in a further embodiment the invention provides a process for the
treatment of inflammatory diseases ATIE 5 i ii i.~i*li sle--cof animals which process comprises
administering to said animals an effective amount of a compound of the invention as hereinbefore
defined. The process may also be used to treat humans.
The compounds of the invention have proved particularly effective in alleviating the symptons of
inflammatory diseases such as rheumatic disease, arthritis and rheumatoid arthritis when applied
topically.

Preferably the compounds of the invention are applied io in the form of a composition comprising a
compound of the invention in admixture with a pharmaceutically acceptable carrier.

The compositions are preferably suitable for topical application to animals to be treated and therefore
may be in the form of a gel, an ointment, a paste, a cream or a lotion.

Compositions comprising a compound of the invention in solution are preferred as they are more
efficient in perfusing the skin.

It is possible that the ingredients of the composition be separately contained within a single container
such that the carrier and active ingredient only come into contact just prior to use. One example would
be an arrangement of the type used to produce "striped toothpaste" wherein the active ingredient
would be produced as stripes on the carrier.

The compounds of the invention have limited stability in the ps-sence of water and therefore the
compositions comprise a non-aqueous carrier. More preferred compositions L- 6 I- I I I- -*irrmm
comprise a compound of the invention dissolved in a non-aqueous lipophilic carrier. Suitable carriers
include monohydric, dihydric and trihydric alkanols such as, for example: short chain (C1 to C10 and long
chaim C12 to 2.10 6a U11~u~,, ,1 n~C-ii'.--gyn C2 0 alcohols including methanol, ethanol, propanol,
butanol and cetyl alcohol; dihydric alcohols such as diethylene glycol; and polyhydric alcohols such as
glycerol.

Even more preferred compositions which have been shown to have an indefinite shelf life, comprise a
compound of the invention in solution in a liquid carrier comprising an alkanol, glycerol and salicylic acid.

The amount of the compound of the invention employed in the compositions will depend to a large
extent on th;⟨br/⟩ 00 inflammatory condition being treated. However, as a general T-⟨4 my rule the
compositions may comprise from 0.1 to 15% w/v of the copper compounds of the invention and
preferably from 1 to 7% w/v.

As previously indicated the compounds of the invention have proved particularly useful in the alleviation
of the symptoms of inflammatory disease when applied topically to the animal in the form of a
pharmaceutical comrposition as hereinbefore defined. The compounds are believed to be efficacious in
such treatments because of their ready penetration ot the skin. Furthermore, the compounds
arebelieved e offer particular advantages in the treatment of inflammatory diseases of humans because
of the non-toxic nature of the compounds and evidence that the compounds are readily cleared from
the treated animal as indicated by the limited duration of the anti-inflammatory effect of the
compounds.

J. AAVt 9 7 The compositions may comprise, in addition to one or more compounds of the invention,
other pharmaceutically active ingredients including other anti-inflammatory agents and conventional
pharmaceutical excipients known in the art.

The invention is now illustrated by but not limited to the following examples.

EXAMPLE 1 Preparation of Cu[H Sal] 2 C2 H5 OH.

Freshly prepared Cu(OH)2 (0.97 g) was added to a solution of salicylic acid (6.9 g) in anhydrous ethanol
CO -4 ml). The suspension was then refluxed on a water bath until all the copper hydroxide had reacted.
The resultant deepgreen-coloured solution was then filtered and green crystals were deposited on
cooling. These were filtered off, washed with cold alcohol and air dried. Yield 3.2 g (Found: Cu, 16.6; C,
50.2; H, 4.2; Cu C16 H1 607 requires Cu, 16.6;⟨br/⟩ C, 50.1: H, EXAMPLE 2 Preparation of Cu[H Sal] 2 CH 3
OH and Cu[H Sal],.

C3H OH.

Cu [H Sal] 2 CH3OH, analysed as Cu C1 5 1407, and Cu [H Sal] 2 C3H7 OH, analysed as Cu C1 7H1 807
were prepared following the procedure described in Example 1 and substituting for anhydrous ethanol,
anhydrous methanol and anhydrous n-propanol respectively.

8 C CO T-⟨ EXAMPLE 3 This example demonstrates the preparation of a composition comprising Cu[H Sal]
2 C2 H 5 H.

Freshly prepared cupric hydroxide (1.0 g) was added to a solution of salicylic acid (7 g) in anhydrous
ethanol ml). The suspension was heated under reflux unti' all the cupric hydroxide had reacted. The
resultant deep-greencoloured solution was filtered to remove any insoluble polymeric copper salicylate
and glycerol (20 ml) was added to the filtrate.

The deep-green c. 0.1 M Cu [H Sal] 2 C2 HOH solution obtained by this procedure was found to be stable
for a prolonged indefinite period and was used in the tests described in examples 4 and EXAMPLE 4 This
example demonstrates the percutaneous absorption of the compoun. Cu[II Sal] 2 C2 Male Wistar rats
(average weight 250 g) were anaesthetised with diethyl ether and an ea of c 20 sq. cm. was shaven high
on their backs. The Cu[H Sal] C2H5OH composition prepared according to example 3 was then applied to
each rat 20 ml per 3 rats). The rats were then returned to their cages and after two days their urine was
collected and compared with the urine of control, untreated, rats following the procedure below.

Urine (5 ml) was acidified with 2M H2SO 4 and shaken '-4 with ethyl acetate (2 ml). After centrifuging the
ethyl acetate layer was analysed by thin layer chromatography on si lica gel (solvent: benzene, diethyl
ether; glacial acetic acid; methanol; 120:60:18:1 by volume) along with samples of salicylic acid,
salicylu'ic acid, gentisic acid and ethyl salicylate.

The results are shown in Figure 1 which is a diagrammatic representation of a chromatogram run under
the conditions described above with solvent flow from origin line, represented by the points of
application of the samples 1 to 9, in the direction of the arrow shown and wherein:⟨br/⟩ 1 and 9
represent the points of application of a sample of a mixture of salicyluric acid and salicyclic acid (b)
chromatographed for conparative purposes;⟨br/⟩ 2 and 8 represent the points of application of a sample
of the vrine extract of rats treated with the Cu[H Sel]2.

composition;⟨br/⟩ 3 and 7 represent the points of application of a sample of the urine extract of conlrol,
untreated, rats;⟨br/⟩ 4 and 6 represent the points of application of a sample of a mixture of gentisic acid
and :thyl salicylate (d) chrcmatographed for comparative purposes; and represents the point of
application of a sample of the Cu[H Sal!2. C2 ~H composition used to treat the rats.

The chromatogram clearly shows the presence of salicylic acid and gentisic acid in the urine of the Cu[H
Sal] 2 Atreated rats indicating that the composition has perfused the skin of the rats.

EXAMPLE This example demonstrates the effectiveness of topical application of the compound Cu[H Sal]
2 C2 H5 OH in alleviating the symptons of artifically induced inflammation in rats.

Male Wistar rats (average weight 250 g) were anaesthetised with diethyl ether and an area of c. 20 sq.
cm was shaven high on their backs. The rats were then separated into 7 groups and treated with 10ml of
the following solutions over a 6 hour period.
Group 1, the control rats were not treated.

(ii) Group 2 rats were treated with 10 ml of an ethanol/ glycerol solution over 6 hours.

(lii) Group 3 rats were treated with 10 ml of a stock solution 7% w/v of salicylic acid in ethanol/ glycerol
This represents a total possible dose of 700 mg of salicylic acid.

(iv) Groups 4, 5, 6 and 7 were also treated with 10 ml solutions derived from the above stock solution to
which increasing amounts of pure Cu(OH)2 had been added:⟨br/⟩ n 11 ri *i *J T Group No.

4 6 7 Cu(OH) 2 (mg) Cu⟨ 40 154 60 231 120 462 200 770 C HOH (mg)2 5 2H These experimental groups
thus received the same dose of salicylic acid but increasing doses of the action compound Cu(H sal) 2
C2H 5OH.

About 6 hours after the topical treatment of the rats inflammation was induced in a foot of each rat in
each of the groups by the injection of 0.1 ml of 1% saline solution of sodium carrageenan into a foot pad
of each rat. The inflammation of the feet of the rats in each group was then monitored by the swelling of
the feet with a micrometer screw-gauge.

lla rP61*YIICP~dyC~L~I~Up-.~ The results are shown in Figure 2 wherein the curves represent the
increase in paw thickness of the rats with time after the injection of the inflammatory agent. The curves
1 to 7 represent the swelling of the paws of the rats of groups 1 to 7 respectively.

Figure 2 clearly shows the anti-inflammatory effect of the topically applied compound Cu[H Sal]2
C2H5OH.2* 25 1 0 OD EXAMPLE 6 Preparation of Cu[C 6H (SH)COO]2 .C2H5 OH.

This compound was prepared as described in Example 1, bu- a ,bstituting 2-thiobenzoic acid (7.7 g) for
the salicylic acid. Yield 2.9 g. (Found: Cu 15.6; C 45.9; H 3.6.

CuC16H1605S 2 requires Cu 15.4; C 46.2; H EXAMPLE 7 Preparation of Cu[C6H (SeH)COO]2 .C2H5OH.

This compound was prepared as described in Example 1, but substituting2-selenibenzoic acid (10.0g) for
the salicylic acid. Yield 8.1g. (Found: Cu 12.3; C 38.1; H CuC16H 1 605Se 2 requires Cu 12.6; C 37.7; H
EXAMPLE 8 Preparation of Cu [CgH (NH2 )COO] 2 .C2H5 OH.

This compound was prepared as described in Example 1, but substituting 2-aminobenzoic acid (6.9g) for
the salicylic acid.

Yield 5.6g. (Found: Cu 16.4; C 50.6; H CuC 16H1 805 N2 requires Cu 16.8; C 50.3; H 12 EXAMPLE 9
Preparation of Cu2 [C6H .CO2.NH.C4N2H2.NH.C7H0 .C2H 5OH.

This compound was prepared as described in Example 1, but substituting 2- [2-carboxyphenyl] amino-4-
[2-carboxyphenyl] amino-pyrimidine (16.8g) for salicylic acid. Yield 13.3g.

(Found: Cu 24.5: C 46.2; H 3.2. Cu2 C2 0H1805N 4 requires Cu 24.5; C 46.0; H C EXAMPLE ~C Preparation
of Cu 2 [C6H4.CO2.NH.C4N2H 2 .NH.CH 04] This compound was prepared as described in Example 1, Y-4
qd4 but substituting 2- [2-carboxyphenyl] amino-4- [2,3-dicarboxyphenyl] amino-pyrimidine (19.0g) for
the salicylic acid. Yield 14.0g.

(Found: Cu 22.4; C 44.7; H 3.3. Cu2C21H 07N4 requires Cu 22.6; C 44.5; H EXAMPLE 11 Preparation of
Cu2[C6 4.CO2.NH.C N2H2.NH.C8H404].C2H5OH.

This compound was prepared as described in Example 1, but substituting 2-[2-carboxyphenyl] amino-4-
[2,4-dicarboxyphenyl] amino-pyrimidine (19.0g) for the salicylic acid. Yield 15.3g.
(Found: Cu 24.7; C 45.8; H 3.7. Cu2C21H1807N 4 requires Cu 24.5; C 46.0; H -13EXAMPLE 12 Preparation
of Cu[CH 4 CO2 .C4 N 2H 2 C1] 2.C2HOH This compound was prepared as described in Example 1, but
substituting 4-(2-carboxyphenyl)amino-2-chloropyrimidine (11.7g) for the salicylic acid. Yield (Found: Cu
12.5; C 56.5; H 3.9. CuC 2 4H1 805N4 requires Cu 12.7; C 56.9; H SEXAMPLE 130 HCH-CHC Preparation of
Cu[C6H .CO2.NH.C8H 9 2.C2H5OH.

00 v-4 This compound was prepared as described in Example 1, but substituting 2-[2,3-
dimethylphenyllaminobenzoic acid (6.8g) for the salicylic acid. Yield (Found: Cu 11.2; C 64.9; H 5.5. CuC
32H3405N 2 requires Cu 10.9; C 65.1; H -14 L*Lir~l~ypFe--i;'~ ~eylVYylur~ienir),*,:,U,,, THE CLAIMS
DEFINING THE INVENTION ARE AS FOLLOWS:⟨br/⟩ 1. Neutral copper (II) complexes characterized in that
they have the formula Cu[CH 4 (X)COO] 2 ROH wherein ROH represents an alkanol, and wherein X can be
located in any position and is SH, SeH or NHR' where R' is NN N- NH N-H IIN S NH CC N -N N CH NH CHC
COOH COOH COOHCOOH COOH -COOH COOH COOH COOH I ii;⟨br/⟩ 15 r 2. Complexes according to
claim 1 characterized in that the alkanol is a C1 to C6 aliphatic alkanol.

3. Complexes according to claim 1 or claim 2 characterized in that they have the formula ROH X HOR 4. A
method of preparing complexes as claimed in any of claims 1 to 3 characterized in that it comprises
reacting an inorganic copper (II) compound with a sibstituted benzoic acid in the presence of an alkanol.

A method according to claim 4 wherein the reaction is carried out under anhydrous conditions.

6. A method according to claim 4 or claim 5, characterized in that the copper (II) compound is a salt or
cupric hydroxide.

7. A method according to any one of claims 4 to 6, characterized in that the alkanol is a C1 to C6 aliphatic
alkanol.

8. A pharmaceutical composition characterized in that it comprises a neutral copper (II) complex of

formula 16 n vIps~ CU[C6H4 COO] 2 -ROH wherein ROH represents an alkanol, and wherein X can be
located in any position and is OH-, SH SeH, NH2 or NHR' where R' is N N_.

COOH f NI COOH COOH N Cl CH 3 CH 3 and a pharmaceutically acceptable non-aqueous carrier.

L~1ti.~ 17 C- I 9. A composition according to claim 8, in the form of an anhydrous gel, ointment, paste,
cream or lotion for topical application.

A composition according to claim 8 in the form of a solution comprising the copper complex, a molar
excess of a substituted benzoic acid of formula C6 H 4 (X)COOH wherein X is as defined in claim 8, and a
non-aqueous solvent.

11. A composition according to claim 8, characterized in that the carrier comprises an alkanol, glycerol
and salicylic acid.

12. A composition according to any one of claims 8 to 11 characterized in that it contains 0.1 to 15% w/v
of active ingredient.

13. A method of treating inflammatory diseases comprising administering to the patient an effective
amount of a complex as claimed in any one of claims 1 to 3 or a composition as claimed in any one of
claims 8 to 12.

14. Any neutral copper (II) complex, method of preparation thereof or composiion containing said
complex substantially as disclosed in any one Example.
DATED this 28th day of May 1981.

BARRY BOETTCHER: WILLIAM RAYMOND WALKER:⟨br/⟩ MICHAEL WELLESLEY WHITEHOUSE By their

Patent Attorneys GRIFFITH HASSEL FRAZER 18 L 9 CI I D 0 0 0 .0 00 00) ,v--4 0 0) 0 q C 00 I' LL II I
41830M8 (D (D 0 Qc 0 al LL czL~ 0 0 0D C Q o ED i AE 2M 00 J 4 U)00 Fig-2 12 3im Time ft rchIllens-c (0.1
ml 17. sodium carrageenan) Co.

CI4 Time after challenge (0.1 ml 10/. sodium carrageenan)