CA2148256A1 - Compositions and methods suitable for therapeutic and pharmaceutical uses - Google Patents

Abstract

2148256 9409798 PCTABScor01 Inorganic compositions comprising a mixture and/or complex of calcium-containing and sulfate-containing materials are disclosed for treating a variety of diseases, injuries and conditions, including wound healing, pain, itch, inflammation, abnormal cell proliferation, or infections caused by fungal, bacterial, rickettsial or viral agents, and similar conditions. The inorganic compositions are derivable from peat or peat-related substances, and may alternatively be synthetically produced.

Description

WOg4/097g8 PCT/US93/10489 ~ -2 1 ~1 ~ X ~
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1 .

MIXTURES OR COMPLEXES CONTAINING CALCIUM AND SULFATE `~
`,,`'~`' . . .
, 1,,,,~, Technlcal_Field -The present invention relates generally to novel ~-compositions and methods :suitable ~or treatment of disease, ;--injury and other disorders. More specifically, the invention relates to inorga~ic calcium-containing and sulfate~
containing compositions, methods of isolation, methods of synthesis, and pharmaceutical compositions suitable for .
accelerating wound healing, pro~idin~ relief of pain, itch or inflammation, reducing abnormal proliferative cell growth, ;~
or providing anti-fungal, anti-~iral, or anti-bac~erial , s activity. `j~.

,.
Back~round of the _n~ention FS''''"
Clinical use of av~ilable treatments for ~iseases involving epidermal conditions is often limited by toxicity, either sys~emic or lo~aI. For example, methotrexate, while `.
generally effective for treating epidermal conditions when : a~ministered orally,~ i9 rarely admi~istered orally for fear of hepatic or bone marxow toxicity. Topical application of methotrexate ha~ been deemed ineffective~ Similarly, although topical application of 5-fluorouracil may be an effective treatment for psoriasis, it is generally considered i~
to be unacceptably irritating. Steroid therapy, whi'e effective, has so m~ny side effec~s that prolonge~ use is ~. ``
discouraged. ~ Photochemot.herapy with psoralens and 7, ultraviolet light, or PW A (psoralens and W treatment), is ~ ~~
;generally eff.ective~for treatment of epldermal conditions, I ~

W094/0979~ P~T/US93/10489 ~ 2 but it is inconvenient and causes acute side effects, as well as having photomutagenic and photocarcinogenic potentials.
Many of the existing treatments for wound healing and the relief of pain, itch and inflammatory conditions are only moderately effective. Moreover, their clinical use is often limited by toxicity or undesirable side effects.
Considerable research effort has been devoted to elucidating the mechanisms involved with such conditions, but few satiæfactory treatments have been developed. Likewise, most therapies available for treating neoplasms and abnormal proliferative cell growth produce undesirable side effects.
The compositions of the present invention are therefore directed to pharmaceutical preparations and methods for treating a variety of disorders.
Summary of the Invention In one aspect, the present invention provide~ novel calcium-containing and sulfate-containing compositions, and analogs and derivatives thereof, that function medicinally, therapeutically or pharmaceutically in the treatment of variou~ diseases, injuries and conditions. Several mixtures and complexes of calcium-containing and sulfate-containing compositions of the present invention ha~e demonstrated significant therapeutic benefit. A first class of inorganic 2S compositions comprises mixtures of a calcium-containing component and a sul~ate-containing compone~tO Mixtures of calcium sulfate and potassium sulfate are especially prefexred.
A 3econd class of inorganic compositions includes complexe~ of a calcium-containing or potassium-containing component and a sulfate-containing compo~ent. Syngenite and apthitalite are especially preferred complexe~ and may be a~ministered as a mixture with one or more of the abo~e~
mentioned calcium-containing or sulfate-containi~g components. A mlxture of ~yngenite and calcium ~ulfate, for ,~:

. .

WO9~/0979~ C~ 5 6 PCT/US93/1048g ,:.. ,,.i.i 1;'' example, i8 especially preferred therapeutic composition of -the pre~ent invention. The compositions of the present invention may also comprise one or more of the following , elements, which may be present in elemental form, ionic form, ' as a ~alt or chelate, or in any other form: sodium;
magnesium; silicon; sulfur; chlorine; potassium; strontium;
zinc; copper; nickel; and manganese. ~ -The inorganic compositions of the present invention can be i~olated from natural materials, such as peat, using the extraction and purification procedures disclosed herein.
The inorganic compositions may alternatively be produced by combining and/or synthesizing the constituent components~
Novel methods for synthesizing high purity syngenite are also disclo~ed herein. ~;
The inorganic compositions of the present invention have produced therapeutic results in a variety of medicinal, ~ i~
pharmaceutical, and therapeutic applications in warm-blooded animals. Th~se applications may be characterized generally a~ promoting wound healing, treatment of pain, inflammation, `
itch, and inhibition of abnormaI proliferative cell growth. `~
Additionally, the inorganic compositions have demonstrated ~ -anti-fungal, anti-bacterial, anti-rickettsial and anti-viral properties and may also be used in cosmetic preparation~
Various delivery sy~tems may be appropriate ~or admini~tering the compositions of the present invention, depending upon condition and preferred treatment regimen.
Topical delivery 3y~tem~ are e~fective and are generally j preferred for most applications of the pharmaceutical composition of the pre~ent in~tention. Topical formulations may be produced by dissolving or combining the inorganic compo~itions of the preqent i~ention in an agueous or ~`J.'~' ~i' ~ona~ueous carrier. Suitable carriers are well known and are r described below.

Briçf Description af the Dr~winqs .

` .~

WO94/Og79B PCT/US93/10489 c~ æ ~ 4 !
Figure 1 shows an elution profile of a 0.5-30Kd fraction from a peat extract purified by High Performance Li~uid Chromatography (HPLC) according to the methods described herein.
Figure 2 illustrates the X-ray powder diffraction analysis of standard gypsum published by the Joint Committee on Powder Diffraction Standards ("JCPDS") ~ibrary.
Figuxe 3 illustrates the X-ray powder diffraction -~
analysis for syngenite published by the JCPDS Library.
Figure 4 shows an x-ray powder diffraction analysis ~:
spectrum identifying gypsum (CaS04 2H20) in a peat extract sample using X-ray powder diffracti~n analysis. -Figure 5 shows a spectrum identifying gypsum (CaSO4 2H2O) and syngenite ~CaSO4-K2SO4-H2O) in a peat extract sample using X-ray powder diffraction analysis.
Figure 6 depicts a spectrum identifying syngenite (CaSO4 K2SO4 ~2O) and apthitalite (K3Na(S0~)2) in a peat extract sample using X-ray powder diffraction analysis.
Figwre 7 illustrates an X-ray powder diffraction ::-analysis spectrum for syngenite produced synthetically according to the methods described herein. ~ -Detailed Description_of~the In~ention The compositions of the present invention comprise a mixture and/or complex of a calcium-containing or po~assium-containing component and one or more sulfate- '~
containing components in a pharmaceutically acceptable ¦
fonmulation that is effective in the treatment of various diseases, injuries, symptoms or other disorders. All refexences to "components" in this application, in wha~e~er fonm, are understood to include a~sociated, dissociated, ionic, neutral, elemental, ~alt, hydrated, and other fo~ms of the constituents. Thu~, for example, a calcium sulfate component may be prese~t in an associated form as a neutral or ionic species; as part of a larger complex; or in a ....

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W~94/09798 PCT/US93/10489 ~ ~
i 2 ~
-dissociated form in which calcium and sulfate are present as distinct, non-complexed neutral or ionic species. The term "composition" also contemplates mixtures of associated, dissociated and complexed constituents.
The term "mixture," as used herein, connotes a composition wherein the constituent components are present in their associated, dissociated, elemental, ionic, salt, hydrated and other forms. Thus, for example, a composition comprising a mixture of calcium sulfate and another sulfate-containing component, such as potassium sulfate, may comprise calcium sulfate and potassium sulfate typically not physically bonded to one another but rather in neutral or ionic forms and/or partially, substantially or completely dissociated into their respective species. A "mixture" of two components may be substantially or entirely dissociated.
It is anticipated that the precise form~s) of the individual components in a mixture will vary depending, for example, upon the relative quantity of each component, the use of aqueous or non-aqueous carriers, a~d the desired pharmaceutical applications or methods of treatment.
The term "complex," as used herein, connotes a composition wherein individual constituents are associated, i.e., bound to one another covalently or non-covalently as a result of hydrogen bonding or other intra-molecular forces.
Complexes may be prese~t in neutral, ionic, salt, hydrated ox other forms.
A1l reference~ to "calcium sulfate" herein are understood to comprehend calcium sulfate in a non-hydrated form (CaSO4~, as well as in hydrated forms, e.q., CaSO4 ~H20 and CaSO4 2H2O (commonly referred to as gypsum), unless a composition~ ~uch as gyp~um, is referred to specifically.
Suitable mixtures of a calcium-containing component and one or more sulfate-containing components include, for example, mixtures of calcium sulfate with anothe~ sulfate-containing component having a constituent chosen from one or ' --':

W094/097~8 PCT/US93/10489 ~, 5~

more of the following: magnesium, potassium, aluminum, sodium, silicon, sulfur, chlorine, calcium, silicon, strontium, zinc, copper, nickel or manganese. Calcium sulfate is a preferred calcium-containing component.
Preferred sulfate-containing components include: MgSO4, K2SO4, Al2 (S04) 3~ 2CaSO4 MgSO4 K2SO4 2H2, CaSO4 K2so4 H20, 3CaO Al203 CaS04 32H20, CaSO4 Na2SO4, Na2SO4 lOH20 and KqSO4-5CaS04. Mixtures of calcium sulfate with potassium sulfate and/or syngenite (CaSO4-K2SO4-H20) are especially preferred. K3Na(SO4) 2, NaAlSi308, and/or KAlSi308 may also be incorporated in the mixtures. The mixtures are combined in a carrier, as described below, and administered for treatment.
Therapeutically important compositions of the present invention may also comprise a complex of a calcium~
containing or potassium-containing components with one or more sulfate-containing components. Syngenite (Ca~S04 K~SO4 H20) is a preferred complex. Other complexes, ; ~
such a~ K3Na~so4) 2 may also be used. According to e~pecially preferred embodiments, a complex of calcium sulfate with one or more other sulfate-containing components, such as ~yngenite, is administered in a carrier, in the form of a mixture with another sulfate-containing component. The mixture of syngenite and calcium 9ulfate is an especially preferred composition. Mixtures of syngenite with other sulfates, such as MgSO4, K2SO4, Al2(S04)3;
2CaSO4-MgSO4-K2SO4 2H20, 3CaO- A1203CaS~432H20, CaS04'Na2S04, Na2SO4-10H20 and K2SO4-5CaS04, may ~lso be used. K3Na(so4) NaAlSi308 and/or KAlSi308 may also be incorporated in the mixture~
The mixtures and complexes forming the compo itions of the present invention may be deri~ed from natural sources, such a~ peat, or they may be derived synthetically. The biologically active inorganic compositions of the present in~ention were initially disco~ere~ in peat extracts, and W094/09798 PCT/~S93/10489 2 1 4 ~ ~
. ,. . :..:
7 ~`
,'''' ''' considerable data relating to naturally derived peat materials has been collected.
According to one aspect of the present invention, the inorganic preparations comprise an alkaline, aqueous or organic, or mixture thereof, extract of peat prepared according to the methods disclosed herein. The term peat, as used herein, refers generally to microbial degradatlon products of biomass, including peat and peat-related substances such as minerals, coal and coal-derived materials, including leonardite and lignite, and humic and fulvic acid preparations.
Peat extracts are-prepared by extracting peat with aqueous solutions, organic solutions or water-miscible organic solvents at temperatures from below room temperature up to the boiling point of the solvents, but preferably below the boiling point of the solvent. Extraction at room temperature is quite suitable; however, the speed of extraction and total amount of active composition isolated are generally enhanced by carrying out the extraction treatment at elevated temperatures. According to preferred embodiments, purified peat preparations are prepared from Bonaparte peat. Bonaparte peat is hypnum peat obtained from Bonaparte Meadows, a peat bog near Bonaparte Lake, Washington, U.S.A. More specifically, the bog is located approximately 25 miles east of Tonasket, Washington, U.S~A., in Secs. 17, 20 and 29, T. 38 N.j R. 30E, in Eastern Okanogan Cou~ty.
Ba~ic, ~aqueous solvents, particularly those containing alkali metal~, alkaline earth metals and ammonium hydroxides, carbonate~ and bicarbona~esl are preferred for peat extraction. Other organic solvents, or a water-miscible organic sol~ent mixed with an aqueou~ solwent, may also be used. It i9 preferred to use extra~ting solvents having a pH of at least 9, preferably using potassium- and sodium-containing bases. Pota~sium hydroxide (~OH) i~ an especially ''`:

W094J09798 ~ PCT/US9~/10489 ... ......
2 ~ J $

preferred base for peat extraction.
Biologically active factors contained in the peatpreparation are separatèd or removed from the residual solids by customary methods such as filtration, ultrafiltration, centrifugation, and decantation.
Peat preparations are complex mixtures which contain inorganic and organic constituents that may have molecular masses as large as several hundred thousand daltons. Various fractions of purified peat preparations have been found to exhibit biological activity and are referred to herein according ~o their molecular mass or fraction or sample number. For example, 10-30Kd peat preparation refers to a peat preparation comprising constituents having a molecular mass between about 10,000 and 30,000 daltons. Various fractions isolated from peat differ in the profile of their biological activities. A peat extract identified as peak #11 (see Example 3) contains a high level of biological activity. Likewise, samples #44 and #46, described in Example 4, exhibit significa~t biological activity.
Biologically active constituents of fractionated peat preparations were identified as CaS04 2H2O (gypsum), CaSO4 K2SO4 H2O (syngenite) and K3Na(so4) 2 (apthitalite) by X-ray powder diffraction analysis. Each biologically active composition was identlfied by compariso~ of an X-ray powder diffraction spectrum of a fractionated peat extract ~o a standard ~pectrum of the Joint Committee of Powder Diffraction Standards (JCPD5) Library. The standard and experimental X-ray power diffractlon spectra are illu~trated in Figs. 2-6.
Th,~ elemental constituen~s prese~t in a peat extract were identified by qualitati~e analy~is using high re~olution X-ray fluorescence spectromet~y (XRF). The followin~ elemental constituents were identified: sodium;
magnesium; silicon; chlorinej potassium; calcium; stro~ium;
'' ~'~.~ '`
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W0~4/09798 PCT/U~93/1048~ ~
21~S23~
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zinc; copper; nickel; and manganese. It is believed that one or more of these elemental constituents contribute to the biological acti~ity of peat preparations.
The preparations o~ the pr~sent invention have been described above 3pecifically with respect to compositions derived from peat. The compositions of the present invention may also be derived ~rom other sources. High purity calcium ~ulfate and hydrated forms of calcium sulfate, including CaSO4 2H20 (gypsum), CaSO4 ~H20, and the like, are commercially available from a variety of sources. Potassium sulfate tK2SO4) and many of the other sulfate-containing compositions described herein are likewise commercially available. Others of the inorganic complexes disclosed herein may not be commercially available but are available from natural }5 sources. For example, K3Na(SO4)2, also known as apthitalite, i~ not commercially available, but may be obtained as a naturally occurring mineral or from other natural sources, i.e., peat, or it may be produced in the lab according to the protocal of Yanat'eva, O.K., et al., Chem. Abstr. 91 (2)~
7031y (1979). Components such as 2CaSO4 MgSO4-K2SO4 2H20, 3CaO Al203 3CaSO4 32H20, CaSO4-Na2SO4, Na2SO4 10H20, NaAlSi308 and KA~Si308 are not readily commercially available, but they may be obtained as naturally occurring minerals.
Syngenite (CaSO4-~SO4-H20~, also referred to as the double salt of gyp~um, is one of the preferred calcium- and sulfate-containing complexe~, but i~ is not available commercially at high purity levels. Syngenite may be obtained as an occurriIlg mineral or from other natural sources, such as mineral deposits or peat. Applicants are aware of the tw~ following reported synthe~es for syngenite:
Calistru, C., et al., Chem~ ~bs~r~ 106(5):31984k (1986); and Yunu~ova, Z., et al., Chem. Ab~tr. 114~10):84755h (1990), but could not produce high purity syngenite according to the publi~hed method~. Applirants therefore developed the following no~el protocol for synthetic production of syngenite.
Syngenite can be synthesized, very expediently and economically, by mixing an aqueous solution of potassium sulfate with an aqueous solution of calcium sul~ate. A molar excess of potassium sulEate is preferably provided to ~he reaction mixture. According to especially preferred embodiments, a molar access of potassium sulfate of about 3 fold to about lO fold is provided in the reaction mixture.
A detailed protocol for syngenite synthesis is provided in Example 6. That synthetic protocol yielded pure (~90 syngenite.
The preferred method for administration of the compositions of the present invention will vary according to the ~ype and location of the disease, inju~y or condition.
Potentially useful methods of administration include topical application of preparation in an suitable aqueous or non-aqueous carrier, injection of the preparation in a carrier, and oral administration. The preparations may also be administered in a solid form, such as a powder or tablet.
The novel compositions are preferably used topically, but may be used orally or parenterally, either individually or in a pha~maceutically acceptable composition further comprising a pharmaceutically acceptable, and preferably inert, carrier or diluent. The term "pharmaceutically acceptable carriers and diluents," as used hereinj contemplates any carrier or other substance that is combined with the biologically active compositions for u~e in any one of the e~umerated methods of administra~ion.
Suitable aqueous and non-aqueous carriers are well known in the art. In general, any liquid, ~xeam, gel or ~imilar sub~tance that does not appreciably react with the active ingredients and which is non-irritating is sui~able.
In a preferred Pmbodiment mixtures and complexes of the prese~t invention are administered in an aqueous carrier, but various non-aqueous solvents or emulsions may also be used , .

:

W094/0~798 PCT/~S93/10489 l S ~ 5 ~ :

~, as carriers. Suitable carriers include, but are not limited to: 1,2,3,-trihydroxypropanol, triethanolamine, EDT,~and the like. In addition, the preparations may also contain fragrances, colors, self-sterilizing agents, odor controllers and thickeners such as natural gums and/or stabilizers.
The biologically active constituents, e.q., syngenite and/or a calcium sulfate complex, are generally pre~ent in a pharmaceutic preparation in an amount of at least about 0.00001% to about 20~, typically about 0.001~ to 2~, and preferably about 0.01~ to 0.5~ by weight. The concentrations of biologically active constituents such as syngenite and/or the calcium sulfate compl~x may be limited by their solubility in a gi~en pharmaceutical carrier or dilùent. In such a case, the limit of solubility can be the preferred solubility. However, higher percentage~ of biologically active co~s~ituents may be obtained by preparing a slurry, or other mixture wherein not all of the mixture or complex is in solution.
The inorga~ic compositions disclosed herein demonstrate therapeutic utility for a broad range of human and veterinary indications, including: promotion of wound healing; reduction of pain, itch and inflammation; inhibition of abnormal cell proliferation; and infections cau~ed by fungal, bacterial, rickettsial or viral agents. More particularly, as described in the appended examples, the inorganic compositions disclosed herein have been found to be active for the treatment of ~kin disordexs such as psoriasis and eczema, acne, seborrheic kerato~is and actinic : keratosis. They are~very effective in treatlng dermatitis, burns and open wounds and pro~ide pain relief from any number of conditions. The inorganic compositions are also useful i~ the pre~ention and treatment of herpes, con~uncti~itis and athlete's foot, and may be efficacious in the treatment of AIDS.
Moreover, inorganic compositions of the present ` `'~

:`~ 214~2)., i~vencion effectively treat disea f~e f which include multiple drug resis~a~ce, Cy9tic f~rosis, cancers, a~thma, rheumatoid ar~hrltis and other i~lamma~ory disorders. Cancersf for whic~ the inventive compositio~s are el~ecti~e includP j-;
souamous call carcinomas, epi~elial carci~omas, bladder -~
tumorq a~a lung tumors. The compositions cf the present .`~:
irventio~ ar~ also sui~able r~r use ~ cosmeti_ a~plicatio~s.
Ad~in s~ration o~ a therapeutically e~fecci~e ~:
amounc o the comoosi~_ons is orefe~ably beguu at che fi~st ncicac__n o~ ~aln or o~her aisorder, and c_rt-.nued unt~
symptoms disa~oear or ceaCfe ~o re~ond f_O ~reatment. A
nt~.era~euc'~ally ef~3cr''~e amour~tf' fne2ns an amounc off~c-~e ~o allevia; one o- ~ore s~mD~oms, or -educe or amelio-ac_ . one or mor causes of the disease, njury cr discr~er.
T~e ~ollowir.a _~amDles are ~resen~3a _~r traf~ ~e ,~ur~oses only a~d shauld not be construed as ~:-;mi eing r:~e -nvention in any way. Re~ister~d Tr~de ~arks are ~arked wit~ an asterisk r~*

zo ~ r~ r^_ n S~ang~ 5,-acr " (~

O~e aram of ~eac rec~ered ~om Po~aparte Meadaws I~-wa.q sti--ed for ~wo hours at -com te~Deracure with 120 ml a~
6 mM ~OY.. The mlxt~re was ce~ uged and the super~ata~
2~ liGuid was designa~ed "Sta~aard Ext-acc" (SE~
Alter~a~ively, i~ a scaled up process, 1 kilogram of peat may ~e 5ri_red wieh 12 lite~ o~ 6 m~ ~OX, followed by ~iltratio~ -co remove unwante~ solids.
'.:
~ -~
One aram or air dried ~eat was ex~racred by hea~
a~d s.~ ,a ~it~ ~20 ~1 or ~ mM ~OH far 20 mlru~es a~
~oiling. ~he sus~enslon was r^- lte~ed and ~ilt_ate was -ererred ~3 as "Standard Bailed ~x~-act" (SBE). .
33 Alternat ~ely, i kilacram o~ peac was sti=~ed with 12 li~e s ~

',.,~ .

18 2 ~3 ~
`~.,; ;, of 6 mM ~OH for 20 minutes at boiling, followed by centrifugacion to remo~e the solids. ~.

Exam~le 2 ``
Pr~aration Q~ ~r~fi~d_P~ Ccm~o~i~io~
The SBE or SE may be us~d "as is,n but, a purified prepara~ion i9 desirable ~or many purDose~ and was provided using ult~a~ltra~ion technicues. Potassium hydroxide (66.4 g) was added with stirring co 88 kilos o~ BonaDarte oeat -~
(approxlmately ~3 kg dry-weight) suspended i~ 190 liters water. Afte~ 24 hours the s~lid~ were allowed to se~tle. :~
. .
The su~er~a~ant li~uid was se~arated hy àecanting or ,ilt.erir.c. Thls solution cdr-esoo~ds .o SE. U~on lyo~hili2ation, th~ 9 501utio~ has been fou~d to viei~ a~
S a~erage of 0.4 mg/ml solids. The S~ was uit=alilte~e~ , through an Amicon*poly~ul_o~e 30 ~d filter, wh}ch re~alned ~`
s teriai of molecular ma5s greate~ than ;0,Q00 dalto~s . .
(~30~d). The re~zi~ed material (~30gdl tyoically contai~ed a~out 0.2 m~ml solids. The filt_ate, aDou~ 130 liter~
contained ma~erials o~tm.olecular mass ~30~d a~d contai~e~ a~
a~erage of about 0.2 mg~ml qolids. A 2S ll_er ~or~o~ 5r ~his c~0~d solution was ultra~lltere~ through a~other Amicon ~ilter wnic~ ~etained mace~ial3 of molecular mass greate~
than 10~d to si~e 250 mi of a reten~a~e con~ai~in~ a~ ave~age o~ 0.1 ~g~ml solLds.

ExamDle 3 Peat wa~ ex~racted~ purified by ult~-afiltracion, ~0 a~d the~ further purl~ed usin~ HP~C. To pre~are a s~andar~
ex~ract, 65 kilcgrams cl Deat and 2~4 ~ra~s o~ ~OH were g~ir-3d ~ 760~ 1 iters a~ ~a~er. ~te_ 24 ho~-s the soli~9 were al'owed to sectle. ~the su~er~.a~a~c liou~d was -ilter~a ~ o~ decan~ed ca produce~the ~Standard Extrac~." The s~andar~
ex~racc t~as ulc~~filterea ~hrough an hm~cor.*hollow f~lte_ ~ ~ 2~ 2v~
.

car~-idge 30~d tO yield a filtrate ccmDrising 740 liters containing ma~erial o~ molecu~ar mass c30~d.
Althcugn filt~ates ha~e bee~ prepared using -:
di~er~nt size exclu ior methods, all c30~d ~rac~iors h~ e ~een found en~lched ~r materlals ha~i~g de~irable biological propertles. Peat pre~aration~ c~mDri~ir~g the c30~d fractio~
may be fur~he~ ~esolved by processins on a .5~d Amico~ S~iral Wound Cart-ldce, wAic~ re~ai~s material having a mo}ecular .~;.;~..
mass '_cm .--,O~d that is suitaDle lor ~P~C. The HPLC i~;
10fractio~ was obtained by ir.jec~i~g a 250 ul por.~o~ of a .5- ::
30~d ex~~ac~ oreo a 2ec.bman 5 mlc_~n 10 mm x 25 cm C-18 (reversed-~hase) High ~ressur~ hiid Chrcmatcgraphy column. .
A gradien~ or soive~s Degi~ni~g wl~h Methanol (100~) a~G :~
5radUally c~nCing tO e~d with deioni7.ed ~ate~ (100~) was passea t-.-oucn che column ac a flow r~te of '.5 ml/mlr.. ~he ~.
eiuate was 3ca~ed by a UV detector se~ at a wa~e ien~h ..
254 r~.
A f-acrion rererred to aq ~eak ~11 contains a hign .`.
c~nce~~~tlon o~ biologically acc~ve mat~rial. Peak #11 ~.
elu~ed ~rom c to 11 minu~ i~ the HPhC sy~ de~cribed ...
aDove~ Figure ' illustrate~ rhe HPhC result~ a~d identi_ies ...
peak ~11. X-ray powder difCrac~i~n analysi9 ide~t_'leq the .
sigu~fic~n~ c~mDositio~ peak ~11 as ~yp9um ECaSO,-2~0]
and synge~ [CaS0,-~,S0,-~.OI; a~d a~hitalite ~Na(S0;) 21 .
Fi~ures 2 asc 3 illustra~e the x-ray powde_ dif~ractio~
tandard spec_-a for gyp~um a~d synge~l~e, re~pectiYely, p~Dlished by the ~C~DS Library. Ficures 4 a~d 5 illu~rate ~pectra ideu~ryi~g gypqum (Figure 4) a~d bot~ ~ypsum a~d :..
synge~ite (Fisure 5) i~ the peat samDle. Figure 5 . illustrat~s a spec~rlm identifyi~s both sy ~-~ite and ...
apthitali te ' ~ a peak X~l peat samDle.

Ex~mple Al~ na~e_~ Dur~i~a~r_~r~rati~n ;.:~
A~ aoue~us ~olu~ion ~ peac was prepared and -~

21~2.~

allowed to stand un~iltered or a time period su~f~icient for a film to form on the sur~ace, usually at lea~ 1 week. ~e ~:
~ilm was carerully skimmed from the surs~ce and mlxed with water. The re~ulti g film solu~ion was ultraril~ered tr_ougn S a l~d A~icon*sDiral wou~d car~ridge to dryness and the ~lgd fraction wa3 discarded. The solution ~l~d was then ~iltered ::
through an Am~con*s~iral wou~d cart-idge with a nomi~al ~.5~d ;~-~
excl.usion. The retentate was lyophilized to dryne~, ...
recons~i~uted ~ wate~ and called Samnle #44. The ri'trate 10o~ ~.5~d was c~ncen~ ed ~y lyophilizatio~ and called SamDle #46.
Samoles ~44 a~d ~46 yield f-ac~-_ns with ~he same -~
'Y.PLC -et~nti_n times as ~ea~ ~11 f-~m t~.~ S-, ana similar ~`
proporrio~ o- calc-~m sulfate (gypsum) as the majc_ aspec- ~
S~ thei- c;nemical c~mocsic~ 0~9 . Sample ~46, when -~ur~.~ea .~ ;
by ~PLC, elu~ed ~s a slugle peak tha~ como--sed tWO
comoound~, Ot which syngenite was the major ccmoone~t.

ExamDle 5 20A~ c~ n of ~ ~r2~n t~ ~an ~t~
Numerous human ~r~al~ were ca~ducted t~ demonstra~e the u~ility and erfec; veness of treacmen~s usi~g the inor~a~.ic camDositions of the present ~ ~Yention. The inorganic c~Do~itions acm~ ter-d to huma~s patie~s in the -~
25~llowing studies were deri~ed from natural pea~ sourc_s u~le~s otherwise indica~ed. The peat ~repara~ion ;~.~
administe~ed t~ human patient~ was isolated from ~o~aparte -:
peat and puri~led as set forth in Exam~le 2. Unle~s otherwlse i~dlcated, a~ ~queous O.~% ~olution by weight Q~
the 10-30gd ~eat preparation ("Peat Pr~arat~onl~) was a~lied ~p~cally ~hree ti~es a day. Other peat f~-actions, incLuai~g a 3-30~d '_ac_:on o~ standard extract, ~eak ~11 and/or sampleq ~44 an~ X46 were applied to~ically tWO ard ~ccasionally chree time~ daily in a lioui~ car_~er in the ~rials i~.dic3~ed. Acueous and emollie~ pea~ pre~ara~ions -. ., `~"`` 2~ ~2~'~

were admlnistered. ~ ~
1,,. :. ,.
p302'~1~. Pat~ 5 1-.5 Studies were carried ou~ c~ huma~ pa~ients 1-5 /~
having long-standi~5 ~qoria~is a~d in whom conve~tio~al .~;
therapy ga~e o~ly poor-to-moderate cortrol. All other .
~reatmenrS ~ere disc~r~t nued, except for o~e patient wno used Diprol~e~ c-~am on cne elbow for comDarison, and anothe~
patient who c^r.ci~ued h~r usu~l twlce weekl~r UVB t~ea~me~ts.
The 10-30~d Pear P~oaration was a~lied topically twice . :~
daily t~ the in~oived sites. The ~e~ults ar~ shown in Table .;~
1. , - ~ .

~e~ult3 ~ ~ m~enrS wlt~ ~u-~- ~d ~; ?~-.l~E3L~
WEE~ C~ ~EATMENT
1 2 3 4 ; --.
~ NC ' '~ NC - ,~1 NC _NC_- 'tl ~C ' SCALINGO 3 2 0 3 2 0 2 30 i 4 0 1 4~ `
ERY~EMAO 3 2 0 3 2 0 2 30 1 4 0 2 3 '-:
THIC~NESS O 3 20 3 2 0 2 30 2 3 0 2 ~ :

W: Wor~e NC: No change I: Impro~ea -;~
~: ComDete c' e-~ri~g i~ one patient 10-30~d Peat g~e~ara~ion was weil tolerated witho~
irritaticn or ~ai~i~c. Smaller, more recen~ and le~s thic~
pla~ues showea early i~Dro~eme~t, i.e., withi~ two week~
The psor1a~is plaouas were leq~ respongiYe wnen c~ra~ic a~d well e~taDli~hed. ~he imDro~eme~t ln erythema and scale or the psor~a~is ~laoue3 wa3 seen i~ thre~ o- the fi~e pat7 e~s ~ ~"~
and i~Dr~veme~t i~ scales wa~ seen i~ _our o~ the r' Y
patients. O~e pa~e~ had com~lete cleari~.g Q~ all his plaoues excep~ ~r a ~m~ll r~sidual area a~ ~he elbow. The ~-10-30~d pea~ ex~rac~ was eouaL i~ e~ acy tO a pocen~

.-', '-'.
:.
. ~

i`: ` 2 1 '1 $ 2 ~ ~ :

steroid applied topically.

Pru~us/Cu~aneous Da~n - Patient~ 5 Patients 6-il had non-urticarial co~ditions. ~his S category i~ exclusive of those with pruritive ecz~ma or Xeroxis. One patient had po~ sca~etic pru_itus. One had persistenc sc-otal ldiopathic pruritus poorly co~rclled wich to~ical steroids. ~o had chxo~ic ~ruritic ~odularis. One had ge~erQ pru~itug/cutaneous pain secondary tO ne~at-c sarcoma. One had intense pruritu~ over the craft a~d keloidal area resuiti~g from third de~ree bur~s ove~ 40~ of her bod~.
None of ~e Dacie~s except the DOS~ sCa~er i-oa~ien.; were can~rolled with oral H-l and ~ 2 antaaoniscs lS eit~er alone or i~ ccmblr.a~ion ~e.g., Seldane~and Zanta~ a~
Doxep~n*aione). App}ication o~ the followinc _o~lccl aqen~s did no~ produce sac_sracto~y rQsults: Prame~ G-l lotion, Zos~r~x* cate~ory 1 or 2 cort~casteroid c-eams wherQ
approp~ate, a~d non-f~uorinaced cort'cos~eroids on t.~e scrotum.
Upon application or a peat preparation c~mDrisi~g 3 30Kd peat preparation and peak #~1, t~erQ wa immediace ~elier ol the pruritus. The pruri~ic nodularis p~tiQ~tS
reduced their exCo-iaticn to a minimal l~Yel. The OO~t scabet~c pruritus cleared o~er a week, bu~ aGai~ ~~lie~ wa~
~mmediate. The pre~iously u~re~pon~i~e scrotal pru~itus was comDleteLy relieved, hu~ reoulred two or thre~ week~ of treatmè~t to o~tain r~lution. The healed t~.ird degre~ bur~
oatie~ had immediate relief and reouired six to eight ap~iica~ions a day to mair.tain relief. This i~tense ap~ acion was not ~asi~l~ on a c~ti~ued basis and thus brier, ~ulc~ but no~ lastl~g reli~ wa~ obtal~ed in t~is ~ anc~. The he~ati- sarccma ~atient was ~ her last ~wo mon~hs o~ lif e and the contir.uous tch/pai n was so sever~
that rorm~l sleep and daily ~unc io~ina was not possibl~
. ...
-., 21~

18 : ~.

In this case, she had noc responded to the a~ove-noted ora~
agencs or Axsain*cream~ Applicatio~ of the peat ?re~aration prcduced an immedia~e imDrovemeut of both t~,e itc~. and the ~`:
deep ~ur~ing pai~ sensation. She re~uired four a~pii-a~ions -~
daily but no longer dug at her ski~ and wa~ able C3 sleep. :
..,.~ .
V~ a~ U~d/Fsc~ P~t~n.~s_'2-.Huma~ patie~cs 12-14 were t~~a~ed wit~ a ~re~araci-n ccmDr~sing peak #11. In all cases, one pruritus ~:
lQ was relieved im~ediatelyl bu~ the vesicuiar-pus~ular comDoner.t was not cont_~lled. Two o~ the pat~enrs reaui~ud , .
sYs~emlc co_.icos~erci~s and eithe~ Ult-avat~ ~ æmovate~
o~c~enc Lor c-~.croi. The chir- evol~ed _-;o puscular psoriasis and is now using P W A.
.
A~ic ~e a ci~ s - Pa~n~s 1~-20 '~uma~ pati e~ts 15-20 having cnrc~ic a~opic ~-de~ma~itis o~ the Lac were trea~ed wit~ 10-30 ~d ~eat ~reparaci~n. All were ~x~eri~nci~g in~e~se ~n~ uq cl the dr~ facial eczema along with inc_easi~ 'icheni~ication `:~ ~
resul~i~g fro~ the rubbi~g and inIl~mmatio~. One also had ~-:
simllar severity i~ his ha~ds/arms along wit~ exoc. a~ion.
In addition to the emoilie~ts, all were usi~g elthe~ Locoid~
sr ~loco~ cr~am ~wice daily o~ the ~aci~l ecz~a without co~r~}. All we~e showing signs or steroid a~rophy.
The pruritus relief was immediate and the derma~itis cleared over 4-5 days. R ~ Ysio~ a~ several weekq waC obserYed i~ two patients and thre~ were eve~tually mai~tained with once daily applicatic~. One ~a~ient ~::
30 coF~letely cle- red 3~ ac~i~e den~atitiq c~~ conti~ued ;rea~men~ for several mon~hs. Afte~ a chree- ee.~ :~iatu~
treatmen~ was re~umea, but ~ithou~ the s.~me reC_onse. Thi~ -~
~atien~ cGul~ no~ ~ully cont-ol che c~ndi~ on 31ely usins a ComDo5it'0n ccm~rising peak ~11. Twice wee.kl~ Lccoi~ cream :.
~S was reoui-ed along wit~ daily aomlniscratlon o~ pea~ ex~rac_ ' , ;--.`'.

,` ' :
~'` `..~ `'.

W~94~09798 PCT/US93/10489 ~.
21ll3~
j<.;.", I

to control the pruritic dermatitis.

Dental,Application - Patient 21 Patient 21 had four wisdom teeth extracted. Two of the teeth were seriously impacted. The patient was given -' a prescription for hydrocodone (a narcotic analgesic) and relea3ed. The patient was in a great deal of pain and, instead of taking the hydrocodone, he swished a few milliliters of a solution comprising peak #11 around in his mouth. The pain was relieved instantaneously. The patient ,~
repeated the administration at 30-minute intervals for about ~';' two hours. Administration at 2-hour intervals seemed to be ~'~
sufficient thereafter. This patient continued with this schedule for approximately two days and was essentially pain ~-free. ~' Burn - Patient 22 '' Patient 22 3ustained second and third degree burns over most of three fingers on her right hand. The burns were cau~ed by contact with a flame and by ~urning nylon that stuck to her fingers. The Emergency Room doctor di~gnosed the 'burns, cut away the burned nylon and skin, and applied sulfadiaæine. The sulfadiazine was later removed and a 3-30Kd peat preparation was applied. Thé patient dressed the wound at least twice daily with bandages soaked wi~h the 3-30Kd peat preparation. The pain abated almost i~mediately '~,~
upon application of the preparation and the wounds remained ', largely pain-free. Within 3 weeks, the patient's fingers were healed, but still pink. After 2 more weeks, there were no indications of scars or wound marks of any ~ort; the skin of the fingers appeared healthy in all respects.~''''', Pain and Le~ions,- Patient 23 Patient 23 had a lamine~tomy and sub equently ~ ' experienced spasms in his lower back. After eight months, ' ,~-~ 2~ 3 I~

1 ~
: ~
the patlènt was t-eated with a 10-30~d Peat preparatlou i~
a cre~m carriQr and he wa~ immediately ~elieved of ?air~
Patient 23 aiso ha~ a long- lasti~g prcblem with his f3Qc, which was repcrred ~o be jungie rot and wnich cau~ea unDeara~le ltchin5 and open lesions. A~ter a~plying the 10-30~d Puri ie~ Peat Preparation c-eam to his feet, t~.e ~air.
and tchi~g subsided and the ~dor disappeared.

C~~~r~ a~ . ~a . Qn 24 Patiert 24 had su~fered f ~-m c;ronic ar.hricis ~or :~
about twelve years arld COnSU1tQd several doc~o_s a:~d chi.3~ac~rs ove_ r~.ese years. ~atlent 24 was _-ea~ea ~1th ---var~-us oral d-~gs anà njec~-or.~ c_ c-rcisone, _ut -~re o .
these ~re~men~s 2~0vided rQlie~ atienc 2~ a~ d a ~-30~d Peac P_e~arac on to~ically ~o G~ ln~ amed ~^0~ ee and shouider cwic daily. Wlt~A~n 5 days, ~erQ was sign~-~can~ r_lier ~n zll arQ~s. Pa~iQrt 24 was aDlQ tQ
disc_~inue use alte~ about ' O days.

E~ G ~ Q~n~cs 2 ~2 ~atie~t 25 had ecz~ma for a~proxlmately 8 years.
.e had bee~ u~re5~0n5i~e tO other anci-ecze~a thera~y and the ec~ema had ne~er comDle~ely disa~peared nor been erfec__~ely treated. ~.e applied a solutior. ci~mpr_sing 0-30gd ?ea~
Preparatior~ (2mg/ml) diluted wi~h 15 ml water and 15 ~i o~
a 0.004% solution of calcium gluconate (final pH 7.;) tO
Well-e9taDli~3hed SpOt 5 Ol eczema. Skin i~ritation ~bu~ing,, thou~ht ro be caused ~y the calcium al UC021at--, occur--_d ~cr apprcxima~ely 3 0 ml~utes . Applicacior~s were c_~tinued t ~ice daily for ai~out 1 0 days. The affec~ed are~ becæ-.,e cruit- red, however, all le9ions ai5a~peared wi~hin 8 days. The ~eaness disappeared afcer ;re~menc with Licex;~i cream ~r 2 cays, leavlr.g only slight discoloracion o~ the skir~. Nc) 1 e~ions have reapDeared ir~ the~ s~Lme 1 oca~ioD.s, bu~ the ec~
~5 con~_nued ~ ~reserl~ itsel~ '3 dif_erent lacarLo~s~ T}~e new ' .-'~,,,"
j ..

W094/09798 214 ~ 2 ;) ~ PCT/US93/10489 L` .
.

ec~ema spots were treated with a peat preparation thatcontained no calcium gluconate. These treatments produced positive results.
Patient 26 applied a 10-30Kd Peat Preparation to eczema covering both legs be~ween his ankles and knees.
Itching was so severe when the patient was in contact with warm or hot water that taking a shower was almost unbearable.
Administering the preparation before a shower greatly reduced itching, used after, the itching stopped within 2 minutes.
Patient 27 had very difficult ec~ema o~er one-third of her body. She treated one arm with a 3-3OKd peat preparation and used the other arm as a control. The treated arm became clear, while the control arm had 25~ coverage of eczèma. Patient 27 also administered t~e preparation on her face and it eliminated the pain associated with the eczema on her face less than 15 seconds after application. Steroids were less effective.
Patient 2~ had mild eczema which could be controlled with steroids using lengthy treatments. Upon application o~ a 3-30Kd peat preparation, he was free from lesions after 7 days.
Patient 29 had a large amount of eczema on her face. A~ter applying a 10-30Kd Peat Preparation twice daily for one week, her face was cleared of all eczema. She continued treatment once daily for anot~er three weeks. The eczema had still not re-emerged after two months.
. .
Wound_Healinq - Patients 30_and_31 Patient 30 applied a 10-30Kd Peat Preparation to sores from abrasions. One of the~e sore~ was infected to the point that it was oozing and weeping. Within 2 to 3 days after treatment with the preparation was ini~iated, the redness and infection was completely gone and complete healing occurred. This healing occurred as ~oon, if not sooner, than another untreated sore which did not have any ... .
:, ,.
,",.,,~.

W0~4/09798 PCT~US93/10489 ~ ~
~7 ~ ~2 ;~

apparent infection. 22 Patient 31 had open and bleeding sores o~ hi~s hands caused by involuntary scratching of eczema during the night.
Cortisone injections controlled the itching for about 4-6 weeks, but the patient was only able to take cortisone shots twice a year. After 2 days o~ treatment with a 10-30Kd Peat Preparation, the itching stopped and healing began. After 7 days, the eczema was completely controlled. When application of the preparation was discontinued, the eczema returned but to a lesser degree. Following 6 days of renewed treatment, the ecæema once again disappeared. Patient 31 continued treatment with the preparation for 8 months with effeckive control of his eczema. No side effects were observed .
Psoriasis - Pa ients 32 and 33 Patient 32 had suffered from psoriasis on his arms i~
and elbows for over ten years. A 1~-30Kd Peat Preparation was applied to one elbow on a twice daily basis for approximately 9 months, with the occasional simultaneous `~
application of fluocinonide cream. Fluocinonide cream alo~e was used on the other elbow. i~
The elbow treated with the fluocinonide alone evidenced only subsided flaking of the skin, but no decrease in the skin lesions. The patient observed significant impr~vement within one week by using the preparation combi~ed with the occasional application of fluocinonide cream. i ~-Flaking and itching had stopped and the lesions on his skin ¦ -were reduced in size. Hair started growing in these areas.
Patient 32 also applied the preparation to open wounds such T
a~ mi~or uts and obQerved good he~ling effects without , `
infection. ! `
Patient 33 had psoriasis that seemed to only - ~ -manifest itself after a strep throat. Her only successful i i~
tre~ment had been with chemotherapeutic agents. Application ;',`

W094/09798 PCT/US93/104B9 ~-~

4 ~

of a 10-30Kd Peat Preparation cleared up the treated psoriatic area in ~ to 3 weeks.

Epidermal Conditions - Patients 34-44 ~uring the winter months, Patient 34 had an extreme case of dry skin and red rash on the inside of her legs. The itching immediately cea~ed upon application of the 10-30Kd Peat Preparation. Within one week, the red rash was gone and ;
the dry skin was completely normal.
Patient 35 applied a 10-30Kd Peat Preparation to treat 9umac poisoning on his legs and arms. He had previously used 1~ cortisone treatment to soothe the burning and itching and to clear up the blis~ers, which took a week to 10 days. The skin would also turn red and peel like a sunburn before the irritation would s~op. After applying the preparation, he had immediate relief from the burning and itching. Within 24 hours th~ blister~ were gone, and within 48 hours the redness was gone and he skin looked normal.
Patlent 36 applied a 10-30Kd Peat Preparation to her lip at the first sign of a cold sore. The preparation stopped the lesion from appeaxing. There was no pain after the fixst application.
Patient 37 experienced mouth sores from overuse of ibuprofen. A 10-30Kd Peat Preparation was applied directly to the sores and cured the co~dition in 12 hours. The healing process generally took 3-5 day~ if untreated.
Patie~t 37 al90 applied the preparation to numerous cuts and ~ --abrasions to effectively avoid infection and accelerate the healing process. Pain was generally controlled within five ~econd~ after application.
Patient 38 had boil-like condition~ as a result of a lingering staphylococcal infection. The condition generally resulted in an infection that required lancing. i ~;-After application of a 10-30Kd Peat Preparation to the affected areas three or four times daily for fi~e days, the ,.:

.

~Q~-7& PCT/US9~/10489 condition completely healed. Patient 38 also applied the preparation to skin blemishes with excellent healing ~esults.
Patient 39 treated a third-degree kitchen stove burn with a 10-30Kd Peat Preparation 10 minutes after the burn was sustained. The associated pain diminished in 15 seconds.
Patient 40 applied a 10-30Kd Peat Preparation to her leg about 24 hours after it was burned. At the time she applied the preparation, the burn was quite painful and blistered. Immediately after treatment, the pain subsided.
Within 24 hours, the blistering was gone and the burned skin was smooth.
Two hours after Patient 41 burned his finger, it , . -was blistered and weeping. After applying a 10-30Kd Peat - ~-Preparation, he experienced immediate relief. The blistering was gone overnight. Patient 41 also applied the preparation to finger inflamed by a steel sliver and ~here was an ~-immediate reduction in pain and pressure. ~-~
Patient 42 experienced itching in his right eye. -~-A~ter one day, his eye became red and inflamed. Flushing , `-with eye wash did ~ot provide relief. After se~eral days, `
the eye was completely stuck closed, very swollen and red, and the patient was diagnosed with conjuncti~itis. The patient applied a cotto~ pad soaked with a 10-30Kd Peat l-Preparation. On the following day, the patient had Yery , ~
little swelling and no pain in his eye, but the eye was still red. He again applied a cotton pad soaked with the . i;
preparation be~ore going to bed. On the follow~ng day, his eye had no swelling, no pain in the eye and no itching. Two days later, the eye was completely healed. -"
Patient 43 applied a ~0-30Kd Peat Preparation to ~ `~
portions of a badly skinned knee. Within two days, all of the sorenes~ and redness was gone and a thin layer scab hadr, .~ ", formed. The untreated area was still sore to the touch. Thei`
scab that formed on the treated area was much thinner than ` ~ ~
:

W094/0979~ PCT/US93/~0489 ~-2~ ~825~` i i that o~ the untreated. There was no pus-like substance at any time on the treated area, but there wa~ a cqntinuing ¦
secretion of pus for five days on the untreated area. The subject reported that overal~ healing of ~he treated wound ~ ;
was at least twice as fas~ as usual.
Patient 44 had aone congolbata on his back, buttocks, and legs. This is a severe, pain~ul condition of ;~
boils which must be frequently lanced. He was being treated with Prednisone at a dose of 27 mg a day. This treatment -~
barely contained the boils. The patient was visiting the ;-Emergency Room at the hospital as much as once a week for -:
lancing. He began using 60 mgs/250 ml~peat preparation #44 in a cream. He was able to reduce the dose of Prednisone to 5 mg a day, with continued reduction of the Prednisone dose thereafter. With continuing treatment, the boils stopped , .. ....
erupting and the pain was diminished.

Funqal Condition - Patient 45 Patient 45 suffered from a chronic athlete's foot lnfection. He had been told by physicians that his condition `-;
was incurable. After one application of a 10-30Kd Peat ~ ~
..........
Preparation, his conditlon began to clear. After the second and third applications, there was no e~idence of the ~ungal infection.
5hinqles - Patient 46 Two children were diagno~ed wi~h shingles. A ~ ~-prescribed medication was used on them for three weeks with ~ -no relief. Peat preparation #44 wa3 administered topically ~ -and provided immediate relief form the pain. In two days, ~ `
the lesions were gone.

Example 6 ~-Mixtures and Com~lexes Inorganic composition mixtures and/or composed of '' ,~' '"'':' W094/~9798 PCT/~S93/10489 2 i ~ , ?. 5 i3 complexes of the present invention were also synthesized and ! -administered to human patients. Inorganic mixtures were prepared using a combination of gypsum (CaS04 2H20) and 3 .
potassium sulfate ~K2SO4). More specifically, a "Mixture" ~ -~
containing O.6 mg of equimolar CaS04 2H20 and K~SO4 in a carrier comprising 2ml of ethanamine-N-N-diethyl- - -trifluoroacetate (EDT) and glycerol in a 1:1 ratio was formulated.
Patient~ 47 and 48 applied the Mixture to small, 0.5 - l.Ocm2 acid burns. The Mixture totally relieved pain within 3-7 minutes after a single application. Treatment -with a water placebo on some sites yielded no pain relief.
Treatment with a 3-3OKd peat preparation yielded temporary ;
pain relief after about 4 minutes, but additional treatments ~- -were required to sustain pain relief. i~
Patient 4~ applied the Mixture topically to treat `
long-term low back pain due to nerve damage caused by disc deterioration. Pain relief occurred within seconds and was sustained. ~
-Example 7 --Synthesis of Synqenite ~-Syngenite was synthesized according to the `
following protocol. Solution A was formulated by dissol~ing ~-125 mmoles of ~S04 in distilled water (450 ml) at room ` -temperature. Solution B was formulated by mixing 2.5 mmoles ~
CaS04 in distilled water ~50 ml) at room temperature with ` `
constant stirring. Solution A was slowly poured into solution B with constant stirring. The reaction mix~ure was --maintained for 4 hr at an isotherm of 38C. ~-Upon evaporation of water, crystals formed which were ~iltered through a membrane filter. Crystals were wa~hed with small amount of ice-cold water:methanol (1:1) then with ice-cold water and dried. The crystals thus ~-~
obtained were recrystalli~ed with ~ater, re~ulting in the ~
: .
:

W094~0~798 PCT/US93/10489 2 1 ~ ~ 2 3 ,`) ~

::
formation of pure (>90~) syngenite.

Example 8 :~
Comparative Studies ~ .
Experiments were conducted to compare the .. -effectiveness of samples #44 and #46 to peak ~11. Based upon .-:
the reports of the test subject, samples #44 and #46 both , -~
worked about 30~ as well as peak #11. When samples #44 and .:~
#46 were administered in an EDT carrier, however, each sample ~;
produced results that were comparable to those obtained with ~;
peak #11. .~
Another patient topically administered three ^- -different preparations to an area of pain. The three ~-preparations included: (1) a 3-30Kd peat preparation; ~2) a .. ;
peak ~11 preparation in a cream; and (3) an equimolar mixture .. :.
of CaSO~ 2H20 and K2SO4 (each about 0.03~ by weight) in 1,2,3-trihydroxypropanol. :The preparations were applied to :`
different areas to reduce intense pain associated with a ..
chronic arthritic condition of many years' duration. .i The patient reported that the 3-3OKd peat ...
preparation produced noticeable but short-lived relief for 20 minutes, with some relief lasting for approximately one hour. He also reported that the peak #}1 preparation ~
provided substantial relief for 20 minutes after application, .`coupled with reduced relief for 3 to 4 hours. The ~-patien~ furthermore reported that the use of the mixture in . ,~ . . .
the 1,2,3-trihydroxypropa~ol carrier produced relief equivalent to the peak #11, and had the additional advantage that it was nèither greasy nor stic~y. ;.
. ;
Examp~e 9 S; ~:
Additional Comparative S~udies s Three human subjects, two sufferi~g low back pain ,~-: and one with pain due to peripheral neuritis of the lower ` ,~
extremities, were each given four blinded samples for uniform :
'~'`.
.

`: ` `::

21~1325~3 i' ,.~..-.
28 :-~

dose topical aoplicatlon. They were i~scruc~ed tO ~first apply sam~le ~1 and then five minutes later to record the level of pain _elief accordins to ~he followi~g scaie: No Relisf=0~, Minor Relief=33~, Significa~t Relief=66~, and . :~
Co3~let~ Relief=100%. I~ No Relief or Minor Reller were ~oted, subjec~s were i~trlcteà to a~ply qamDle #2 to a ~ew ~.;:.topical area of pain. The 9ame ge~eral scheme was ccn~i~ued n~il all fou- 9am~l es had bee~ testec.. The results 0!- the test were as follows: .

Su~iec~s' ~es~onse~

sam~iel ~ ~
~o. ¦com~le~e Isi~ can~ IMi~orlRelier l~ura-ion l ~ ;
~ ~ .;~
lS Xl 1 3/3 1 I I!1-4- :~~s I
_. _ . . _ _ _ _ X2 1 1/3 2/3 3 hours ~3 1 _ I 1/3 2/3--~ ~ ,~
~4 ~ /3 1 ~ `;i-`
~ ,`: .
The sm~ll size of t~is group perm~ts oniy one clear concluqic~. Sampie #1 ~rovide~ clear a~alge~ic 3ene~i ~ior all 5UDj ec~s compared to ~he ocher samDles. The comDosit o~
o~ samole #l was 0.25~ CaS0~-2~0 Dlus 0.5~ syngenit~ ir UNIBAS~ cr3am. UNIBASr is a commerc~ally available ~ooical 2S cream. The compositio~ or gamDle5 ~2, #3, and ~4, reqpeccively, was a~ follow~: ~2 = U~I3A5~ alo~e, ~3 = 0.25%
syngeni~Q in UNI~ASE*~nd ~4~ .25% CaSO~-R~S0~-~.0 i~ UNIBAS~
I~ the subject with pain due to neuritis there ~ also a ~ery sicnif ca~c le~el o~ lacal ~flamma~ion which was ~ir~ally ccmpletely ameliorated by ~he aDplicacior- cr s~mDle #1.
~his exam~le demons~r~tQs se~eral i.~Dcr~an_ ~spec~s af t~Q irl~encion. ~irs~, t utili~ed syngenice pre_~rQd by the claimed ne~ syncheti_ proce~s. Second, i~ show2a tha~
a 'or~ula~lon comoo~ed o. calc~lm sulCa~o and anocher wO ~4/Og798 2 1 ~1 r~ PCT/US~3/10489 ,r 1~ ' sulfate-containing compound (syngenite) is responsible for the amelioration of pain and inflammation in human s~ubjects.
Third, since both of these compounds are known to be part of ' `~
the 3-30Kd, 10-30Kd and #11, #44 and #46 fractions in i ~ ;-variable proportions, we can conclude that at least some of the beneficial results produced by these preparations are due to the presence of these components. ~

Example 10 -Veterinary Applications The inorganic compositions of the present invention also demonstrate significant utility ~or veterinary -applications. The results of several animal experiments are ., .
present below.
Animal subject A was a dog that started intensive scratching after a ~isit to the seashore. After two to five days~ itching increased with some hair loss as a result of continued scratching. On day six, the dog was sprayed with flea and tick powder but showed no improvement. The bites continued to worsen and soon secreted a pu~-like substance.
The dog was treated by applying a 10-30Kd Peat Preparation in spray form. The spray was re-applied 8 hours later. On the following day, there was no pus and less scratching. The preparation was reapplied three times during the day and the bites were smaller and less red. After one more day of treatment, the bites were smaller in size, only pink in color and there was no scratching.
Animal subject B was a race horse uffering from sores on its legs. The horse wa~ diag~osed as being allergic to mud and was treated witn triamcinolone 0~1~ acetonide cream. He had scabbing and oozing of pus on his legs and was sore and stiff. There was no improveme~t after four months of treatment under ~eterinary supervision. A 10-30Kd Peat Preparation was prayed directly on ~he hor~e' 9 legs o~ce a day. After three days, healing was noted. After o~e week, WQ94/0979g P~T/US93/10489 p ~
æl l~2~

the infected areas were healed and the horse's hair was growing back. ~
Animal subject C was a large dog that had serious eczema and dry skin since birthO Numerous sprays, powders, shampoos, pills and injections were administered without success. After spraying the dog with a 10-30Kd Peat Preparation for two weeks, itching was resolved and there was less flaking and dry skin.
Animal subjects D and E were adult Labrador Retrievers infested with fleas. A 10-30Kd Peat Preparation was applied in a liquid form three to four times daily to the thighs, perianal region and tails of two dogs. No other attempt was made to treat the ~leas on the dogs or in their environment. Prior to treatment, bitten fur and a flea-induced denmatitis was obvious. Immediate reduction inbiting, scratching and licking of the in~olved sites was apparent upon initial application of the preparation. Since the flea population was not reduced, four daily treatments were xequired for control of the pruritus and clearance of the dermatitis.
Animal subject F, a German Short Hair dog, was treated with the 10-30Kd Peat Preparation for a non-flea associated, non-specific dermatitis. ~pplications were inconsistent and usually twice daily. There was a noticeable reduction in the scratching and dermatitis within one week.
An open clinical trial was conducted in an attempt to determine the efficacy of a 10-30Kd Peat Preparation when topically applied for treating a variety o~ itching canine skin disea~es. Most of the dogs suffered from allergy~
as~ociated itch (flea bite, inhaled and/or food allergies~.
In over 50 dogs evaluated, the prepara~ion was instrumen~al in the relief of itch in greater than 50~ of the subjects.
No adver~e side effects were noted. Itch relief was usually noted within three days of initiating therapy. Additionally, the preparation demon~trated significant anti-inflammatory -.,''`~",~-WO 94/~97~8 ~ 2 ~ ~ PCT/US93/10489 ~ ~

.,.. , , 1": ' ~`:

properties ~rapid decrease in erythema and swelling) when ¦, applied to allergy-a~sociated rashes. Prepa~rations ~ ~
comprising syngenite produced synthetically according to the ! `::
protocol given in Example 7 produced similar therapeutic ,~
results when administered to dogs.

Example 11 !;
Treatment of Bovine Squamous Cell Carcinoma Bovine ocular squamous cel~ carcinoma of cattle is a common neoplasm of the bovine eye and adenexa. Squamous cell carci~omas may also affect other species of animals.
. .
Bovine ocular squamous cell carcinoma is best known by its colloquial name of Cancer-eye. The initial lesion may be on the eyelid or any structure in the conjunctival sac except the vascular cornea or the pigmented eyelid. The lesion develops through three ~tages~ The first stage is the formation of a plaque; the second s~age is formation of a papilloma; and the third stage is the squamous cell carcinoma.
The first two stages are no~-malignant and have up to an 88% regre~sion rate. The third stage i9 malignant and does not regre~s. These carcinoma3 develop most commonly on the nictating membrane, the eyelids, and the corneal limbus.
They grow rapidly and are actively invasive, often metastasizing to the lymph nodes. The ab~ve-described cancer most usually affects Hereford cattle, but has been found in Ayrshire, Simmental, Shorthorn, Hol~tein and ~ross-bred ani~als thereof. Several cattle were successfully treated with a 10-30Kd Peat Preparation.
, . ...
A cow having bovine ocular ~q~amous cell carcinoma was treated by injecting 5 ml of a solution containing 3 mg/ml 10-30Kd Peat Preparation into the sarcoma. Ten days later the ti99ue firmed up, blood supply increased and the tumsr had shrunk con~iderably. Three week~ later, there were no ~isible sign~ of the tumor remaining.

.' ~'`~' `

W~94/09798 PCT/US9~/10489 % ~4~ i 32 Another cow was treated using a slightly different treatment regimen. The original lesion measured~ 5 cm x 2~ cm. Initial treatment consisted of debriding the necrotic areas, and then suturing 4 gauze pads over the right eye S after injecting 1 ml of a 10-30Kd Peat Preparation into the tumor. The gauze was subsequently soaked liberally with the preparation. The soaked gauze was covered and an eye patch was applied. This treatment procedure was repeated weekly for five weeks. During this time, the size of the tumor reduced substantially.
A 12-year old purebred Hereford cow had a s~uamous cell carcinoma on its upper and lower left eyelids and nictating membrane. Necrotic tissue was debrided under local anesthesia, a biopsy sample was taken, and 3 ml of the 10-3OKd Peat Preparation was injection into the lesion. The examination and treatment were repeated after 9 days and considerable improvement was noted~ The treatment was repeated after 49 days. A second growth was found on the right eye, which was also treated. After two months, the cow appeared to be healthy and tumor-free.
Another cow had a ~uamous cell carcinoma lesion .5 cm2 on its corneal limbus. The growth was surgically removed, and then 0.5 ml of 10-30Kd Peat Preparation was injected into area. No further treatment was administered for one year, when a small plaque was removed.
The examples presented abo~e are to be considered in all respects as illustrative and not restrictive. The scope of the invention is indicated by the appended claims only, and all modi~ications which come within the meaning and equivalency of the claims ~herefore are intended to be embr~ced therein.

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Claims (42)

CLAIMS:

1. A pharmaceutically acceptable composition comprising a complex of calcium-containing component, a potassium-containing component and a sulfate -containing component in a pharmaceutically acceptable carrier or diluent, the calcium-containing component, the potassium-containing component and the sulfate-containing component being bound to one another covalently or non-covalently.

2. A pharmaceutically acceptable composition according to claim 1, comprising a complex of a potassium-containing component and calcium sulfate.

3. A pharmaceutically acceptable composition according to claim 1, comprising a complex of a calcium-containing component and potassium sulfate.

4. A pharmaceutically acceptable composition according to claim 3, wherein the calcium-containing component is calcium sulfate.

5. A pharmaceutically acceptable composition according to claim 4, wherein the complex of calcium sulfate and potassium sulfate is syngenite.

6. A pharmaceutically acceptable composition according to claim 1, comprising a calcium-containing component in addition to the complex.

7. A pharmaceutically acceptable composition according to claim 1, additionally comprising a complex of potassium-sodium-sulfate.

8. A pharmaceutically acceptable composition according to claim 7, wherein he complex of potassium-sodium-sulfate is apthitalite (K3Na(SO4)2).

9. A pharmaceutically acceptable composition according to claim 1, additionally comprising one or more of the following components: MgSO4; K2SO4; Al2(SO4)3; CaSO4;

CaSO4?1/2H2O; CaSO4?2H2O; 2CaSO4?MgSO4?K2SO4?2H2O;
3CaO?Al2O3?3CaSO4?32H2O; CaSO4?Na2SO4; Na2SO4?10H2O;
K2SO4?5CaSO4; NaAlSi3O8; and KAlSi3O8.

10. A pharmaceutically acceptable composition according to claim 1, additionally comprising one or more of the following constituents: sodium; magnesium; silicon;
chlorine; potassium; strontium; zinc; copper; aluminum;
nickel; and magnanese.

11. A pharmaceutically acceptable composition according to claim 1, wherein the carrier of diluent is aqueous.

12. A pharmaceutically acceptable composition according to claim 1, wherein the carrier or diluent comprises 1,2,3-trihydroxypropanol.

13. A pharmaceutically acceptable composition according to claim 1, wherein the carrier or diluent is a topical cream.

14. A pharmaceutically acceptable composition according to claim 1, wherein the complex is derived from peat.

15. A pharmaceutically acceptable composition according to claim 14, wherein the complex is derived from a .5-30 Kd peat fraction.

16. A pharmaceutically acceptable composition according to claim 14, wherein the complex is derived from a 10-30 Kd peat fraction.

17. A pharmaceutically acceptable composition comprising a mixture of a calcium-containing component, a potassium-containing component and a sulfate-containing component in a pharmaceutically acceptable carrier or diluent.

18. A pharmaceutically acceptable composition according to claim 17, wherein the calcium-containing component is a calcium sulfate.

19. A pharmaceutically acceptable composition according to claim 17, comprising a mixture of a calcium-containing component and potassium sulfate.

20. A pharmaceutically acceptable composition according to claim 19, wherein the calcium-containing component and calcium sulfate.

21. A pharmaceutically acceptable composition according to claim 17, wherein the sulfate-containing component and syngenite.

22. A pharmaceutically acceptable composition according to claim 17, comprising a complex of a calcium-containing component and a sulfate-containing component in addition to the mixture, the calcium-containing component and the sulfate-containing component being bound to one another covalently or non-covalently.

23. A pharmaceutically acceptable composition according to claim 17, additionally comprising a complex of potassium-sodium-sulfate, the potassium, sodium and sulfate being bound to one another covalently of non-covalently.

24. A pharmaceutically acceptable composition according to claim 23, wherein he complex of potassium-sodium-sulfate is apthitalite (K3Na(SO4)2).

25. A pharmaceutically acceptable composition according to claim 17, additionally comprising one or more of the following components: MgSO4; K2SO4; Al2(SO4)3; CaSO4;
CaSO4?1/2H2O; CaSO4?2H2O; 2CaSO4?MgSO4?K2SO4?2H2O;
3CaO?Al2O3?3CaSO4?32H2O; CaSO4?Na2SO4; Na2SO4?10H2O;
K2SO4?5CaSO4; NaAlSi3O8; and KAlSi3O8.

26. A pharmaceutically acceptable composition according to claim 1, additionally comprising one or more of the following constituents: sodium; magnesium; silicon;
chlorine; potassium; strontium; zinc; copper; aluminum;
nickel; and magnanese.

27. A pharmaceutically acceptable composition according to claim 17, wherein at least one of the components is derived from peat.

28. A pharmaceutically acceptable composition according to claim 27, wherein at least one of the components is derived from a .5-30 Kd peat fraction.

29. A pharmaceutically acceptable composition according to claim 27, wherein at least one of the components is derived from a 10-30 Kd peat fraction.

30. A method for synthesizing a complex of calcium sulfate and potassium sulfate comprising: mixing an aqueous solution of calcium sulfate with an aqueous solution and molar excess of potassium sulfate.

31. A method for synthesizing a complex according to claim 30, wherein a molar excess of potassium sulfate of about 3-fold to about 10-fold is provided in the reaction mixture.

32. A pharmaceutically acceptable composition comprising a complex of calcium sulfate and potassium sulfate produced according to the method of claim 30, the complex having a purity of more than 90%.

33. A method for reducing inflammation in warm-blooded animals by administering he pharmaceutically acceptable composition of one or claims 1, 17 or 32.

34. A method for reducing pain in warm-blooded animals by administering the pharmaceutically acceptable composition of one of claims 1, 17 or 32.

35. A method for reducing itch in warm-blooded animals by administering the pharmaceutically acceptable composition of one of claims 1, 17 or 32.

36. A method for reducing abnormal proliferative cell growth in warm-blooded animals by administering the pharmaceutically acceptable composition of one of claims 1, 17 or 32.

37. A method for promoting wound healing in warm-blooded animals by administering the pharmaceutically acceptable composition of one of claims 1, 17 and 32.

38. A method for treating fungal infections in warm-blooded animals by administering the pharmaceutically acceptable composition of one of claims 1, 17 or 32.

39. A method for treating bacterial infections in warm-blooded animals by administering the pharmaceutically acceptable composition of one of claims 1, 17 or 32.

40. A method for treating viral infections in warm-blooded animals by administering the pharmaceutically acceptable composition of one of claims 1, 17 or 32.

41. A cosmetic preparation comprising a complex of a calcium-containing component, a potassium-containing component and a sulfate-containing component.

42. A cosmetic preparation comprising a mixture of a calcium-containing component, a potassium-containing component and a sulfate-containing component.