United States Patent [191
`Ueda et a1.
`
`[11] Patent Number:
`[45] Date of Patent:
`
`4,916,138
`Apr. 10, 1990
`
`[54] SOLID DISPERSION COMPOSITION OF
`FR-900506 SUBSTANCE
`[75] Inventors: Yoshio Ueda, Kobe; Fumio Shimojo,
`Kawanishi; Yasuo Shimazaki,
`Takarazuka; Kazutake Kado, Ikeda;
`Toshiyasu Honbo, Kobe, all of Japan
`Fujisawa Pharmaceutical Co., Ltd.,
`Osaka, Japan
`[21] Appl. No.: 224,235
`[22] . Filed:
`Jul. 25, 1988
`
`[73] Assignee:
`
`[63]
`
`Related US. Application Data
`Continuation of Ser. No. 32,572, Apr. 1, 1987, aban
`doned.
`Foreign Application Priority Data
`[30]
`Apr. 2, 1986 [GB] United Kingdom ............... .. 8608080
`
`[51] Int. C1.4 ................................ ..; ......... .. A61K 31/44
`[52] US. Cl. .................................................. .. 514/294
`[58] Field of Search ....................... .. 514/294; 374/294
`[56]
`References Cited
`U.S. PATENT DOCUMENTS
`
`3,852,421 12/1974 Koyanagi et a1. .
`4,259,314 3/1981 Lowey .
`4,265,875 5/1981 Byrne et a1. .
`
`4,369,172 1/ 1983 Schor et a1. .
`4,389,393 6/1983 Schor et a1. .
`4,412,986 11/1983 Kawata et al. .
`4,654,206 3/1987 Okuda et a1. .
`
`FOREIGN PATENT DOCUMENTS
`
`0184162 6/1986 European Pat. Off. .......... .. 514/294
`852904 4/1986 Greece .
`2050828 1/ 1981 United Kingdom .
`2053681 2/1981 United Kingdom .
`2111386 7/1983 United Kingdom .
`2122085 1/1984 United Kingdom .
`
`OTHER PUBLICATIONS
`Pharmaceutica 1981, No. 4, pp. 66-67, Reviews, Tent
`sova et a1, “Solid Disperse Systems in Pharmaceutics”.
`Primary Examiner-Jerome D. Goldberg
`Attorney, Agent, or Firm-—-Oblon, Spivak, McClelland,
`Maier & Neustadt
`ABSTRACT
`[57]
`This invention relates to a solid dispersion composition
`comprising FR-900506 substance and water-soluble
`hydroxypropyl methylcellulose which is capable of
`dispersing the FR-900506 substance, the FR-900506
`substance and hydroxypropyl methylcellulose being in
`the ratio of 1:0:1 to 1:20 by weight.
`
`3 Claims, N0 Drawings
`
`NOVARTIS EXHIBIT 2037
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`Page 1 of 5
`
`

`
`1
`
`SOLID DISPERSION COMPOSITION OF
`FR-900506 SUBSTANCE
`
`This application is a continuation of application Ser.
`No. 32,572, ?led on Apr. 1, 1987, now abandoned.
`The present invention relates to a solid dispersion
`composition comprising l7-allyl-l,14-dihydr0xy-l2-
`[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]
`23,25-dimethoxy- l 3, 19,21,27-tetramethyl-1 1,28-dioxa
`4azatricyclo[22.3. 1 .04'9]octacos- 18-ene-2,3, l0, 1 6-tetr
`aone (hereinafter referred to as FR-900506 substance).
`More particularly, the present invention relates to a
`solid dispersion composition comprising the FR-900506
`substance and a water-soluble polymer.
`The FR-900506 substance used in the present inven
`tion is novel and can be represented by the following
`chemical formula:
`
`15
`
`HO
`
`20
`
`25
`
`CHrCH=CH2
`
`CH3
`
`OCH3 OCH3
`
`30
`
`35
`
`4,916,138
`2
`disadvantage could be overcome by dispersing the FR
`900506 substance with a water-soluble polymer to pre
`pare a solid dispersion Composition, and have com
`pleted the present invention.
`According to the solid dispersion composition of the
`present invention, solubilization of the FR-900506 sub
`stance has been achieved and hence the bioavailability
`of the FR-900506 substance in human body can be dras
`tically improved.
`Further, according to the present invention, the FR
`900506 substance become stable and can be released
`sustainedly, and therefore said substance can be phar
`macologically active for a long time in the body and its
`toxicity can be reduced thereby.
`The solid dispersion composition of the present in
`vention is explained in more detail in the following.
`The solid dispersion composition of the FR-900506
`substance can be prepared by a conventional method,
`for example;
`(1) dissolving the FR-900506 substance in‘an‘organic
`‘solvent, and
`(2) adding a water-soluble polymer to the resultant
`solution, and
`(3) if necessary, suspending the additives such as excipi
`ent, disintergator, and the like, in the resultant suspen
`sion or solution, and then
`(4) removing the organic solvent from the resultant
`homogeneous suspension in a conventional manner.
`And, in case that more homogeneous solid dispersion
`composition is desired, the homogeneous suspension is
`prepared in the above procedure (2) and then subjected
`to the following subsequent procedures.
`(5) dissolving the suspension prepared in the above
`procedure (2) in an organic solvent, and
`(6) if necessary, suspending the additives such as excipi
`ent, disintegrator, and the like, in the resultant homo
`geneous solution, and then
`(7) removing the organic solvent in a conventional man
`ner.
`The organic solvents to be used in the procedure (1)
`are not restrictive and are any solvents which are capa
`ble of dissolving the FR-900506 substance such as alco
`hol (e.g. methanol, ethanol, propanol, isopropyl alco
`hol, etc.), ethyl acetate, diethyl ether, and the like, in
`which the preferable ones may be lower alkanol.
`The water-soluble polymers to be used in the proce
`dure (2) may be a water-soluble cellulose polymer
`which is capable of dispersing the FR-900506 substance,
`such as hydroxypropyl methylcellulose. The hydroxy
`propyl methylcellulose can be used under various vis
`cosity.
`The quantity of the water-soluble polymer is not
`restrictive and is any one, by which the FR-900506
`substance can be dispersed, and suitable quantitative
`ratio of the water-soluble polymer and the FR-900506
`substance by weight may be from 0.1:1 to 20:1, prefera
`bly 0.3:1 to 10:1, more preferably 0.5:1 to 5:1, and the
`most preferably 1:1.
`The additives to be optionally used in the procedures
`(3) and (6) may be a conventional ones used in the field
`of pharmaceutical preparation such as excipient (e.g.
`lactose, sucrose, starch, mannitol, etc;), disintegrator
`(e.g. croscarmellose sodium, carboxymethyl cellulose
`calcium, low substituted hydroxypropyl cellulose, so
`dium starch glycolate, microcrystalline cellulose, etc.),
`and the like, and these excipient and disintegrator can
`be used at the same time or independently.
`
`40
`
`The FR-900506 substance was isolated in a pure form
`from culture broths prepared by fermentation of a FR
`900506 substance-producing strain belonging to genus
`Streptomyces, among which Streptomyces tsukubaensis
`No. 9993 has been newly isolated from a soil sample
`collected at Toyosato-cho, Tsukuba-gun, Ibaraki Pre
`fecture, Japan. And a lyophilized sample of the newly
`isolated Streptomyces tsukubaensis No. 9993 has been
`deposited with the Fermentation Research Institute,
`Agency of Industrial Science and Technology (No. l-3,
`Higashi l-chome, Yatabemachi, Tsukuba-gun, Ibaraki
`Prefecture, Japan) under the deposit number of FERM
`P-7886 (deposited date: Oct. 5th, 1984), and then con
`verted to Budapest Treaty route of the same depository
`on Oct. 19, 1985 under the new deposit number of
`FERM BP-927.
`_
`The FR-900506 substance possesses pharmacological
`activities such as immunosuppressive activity and anti
`microbial activity as described in the published Euro
`pean patent publication No. 184162 (publication date:
`June ll, 1986) and therefore is useful for treatment and
`prevention of rejection by transplantation, graft-versus
`. host diseases by medulla ossium transplantation, auto
`immune diseases, infectious diseases, and the like.
`However, when orally administered, the ratio of the
`absorption of the FR-900506 substance into blood is
`insufficient due to its insolubility into water, and the
`FR-900506 substance has the disadvantage of its poor
`bioavailability in oral administration.
`As a result of an extensive study, the inventors of the
`present invention have discovered the fact that said
`
`55
`
`65
`
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`
`

`
`4,916,138
`3
`The quantity of the additives is not restrictive and
`suitable quantitative ratio of the excipient or disintegra
`tor and the EFL-900506 substance by weight, if used,
`may be from 0.1:1 to 20:1, preferably 0.5:1 to 5:1, and
`more preferably 1:1 to 3:1, respectively.
`The organic solvents to be used in the procedure (5)
`are not restrictive and are any solvents capable of dis
`solving the suspension of the preceding procedure (2),
`such as chloroform, dichloromethane, and the like.
`The solid dispersion composition of the present in
`vention prepared by the above—mentioned procedures
`can be used by itself as a pharmaceutical preparation for
`oral administration and also can be converted into vari
`ous dosage forms such as powders, ?ne granules, gran
`ules, tablets, capsules, injection, and the like, according
`to a conventional manner. If desired, conventional col
`oring agents, sweeting agents, ?avouring agents, dilu
`ents, lubricants, and the like, (e. g. sucrose, lactose,
`starch, crystalline cellulose, synthetic aluminum silicate,
`magnesium stearate, talc, etc.) may be compounded
`with the solid dispersion composition.
`The solid dispersion composition of the FR-900506
`substance and the various preparations thereof prepared
`by optionally converting said solid dispersion composi
`tion into various dosage forms as mentioned above,
`have remarkably improved solubility and absorptive
`ness into blood in comparison to the crystals of the
`FR-900506 substance per se.
`In order to show the usefulness of the solid dispersion
`composition of the present invention, the test results are
`given in the following.
`[I]Dissolution Test:
`
`20
`
`25
`
`Test Samples
`(A) Solid dispersion composition of the FR No. 900506
`substance prepared in Example 2;
`(B) Solid dispersion composition of the FR No. 900506
`substance prepared in Example 4;
`(C) Crystals of the FR No. 900506 substance per se
`prepared in Reference;
`Test Method
`The tests were carried out according to the paddle
`method prescribed in Method 2 of the dissolution test in
`The Pharmacopoeia of Japan (tenth edition) using
`water as test solution and the dissolution rate at 100 rpm
`after the speci?ed minutes from the beginning of each
`dissolution test was measured.
`
`35
`
`45
`
`50
`
`Test Results
`The dissolution rate of the solid dispersion composi
`tion of the FR No. 900506 substance is shown in the
`following table.
`
`55
`
`Test Samples
`
`15 min.
`
`Dissolution Rate (%)
`30 min.
`60 min.
`
`A
`B
`C
`
`54.5
`74.0
`0
`
`74.6
`91.8
`0
`
`88.2
`I00
`5.0
`
`90 min.
`
`93.0
`100
`10.8
`
`[II] Bioavailability Test
`Test Sample
`Solid dispersion composition of the FR No. 900506
`substance prepared in Example 2;
`
`65
`
`4
`Test Method
`The above sample, which contains 10 mg/kg of the
`FR No. 900506 substance, was orally administered to
`several dogs, which had been withheld from any food
`overnight in a crossover design. The plasma concentra
`tion of the FR No. 900506 substance was determined by
`high performance liquid chromatography at l, 2, 4 and
`6 hours after administration.
`
`Test Results
`The plasma concentrations of the FR No. 900506
`substance at each time are shown in the following table.
`
`Time (hours)
`1
`2
`4
`6
`
`Plasma Concentrations (ug/ ml)
`0.44
`0.69
`0.53
`0.45
`
`As clearly seen from the above two test results, the
`solid dispersion composition of the present invention
`possesses good dissolution rate and pattern, and further
`good bioavailability.
`The present invention is explained according to the
`following Examples.
`
`Reference
`Isolation of Streptomyces tsukubaensis No. 9993
`Streptomyces tsukubaensis No. 9993 was isolated by
`using dilution plate techniques as shown in the follow
`ing.
`About one gram soil which was collected at Toyosa
`to-cho, Tsukuba-gun, Ibaraki Prefecture, Japan, was
`added to a sterile test tube and the volume made up to
`5 ml with sterile water. The mixture was then blended
`for 10 second by a tube buzzer and kept on 10 minutes.
`The supernatant was sequentially diluted by 100 fold
`with sterile water. The diluted solution (0.1 ml) was
`spread on Czapek agar supplemented with thiamine
`hydrochloride (saccharose 30 g, sodium nitrate 3 g,
`dipotassium phosphate 1 g, magnesium sulfate 0.5 g,
`potassium chloride 0.5 g, ferrous sulfate 0.01 g, thiamine
`hydrochloride 0.1 g, agar 20 g, tap water 1000 ml; pH
`7.2) in a Petri dish. The growing colonies developed on
`the plates after 21 days incubation at 30° C. were trans
`ferred to slants [yeast-malt extract agar (ISP-medium
`2)], and cultured for 10 days at 30° C. Among of the
`colonies isolated, the Streptomyces tsukubaensis No.
`9993 could be found.
`Fermentation
`A preculture medium (100 ml) containing glycerin
`(1%), corn starch (1%), glucose (0.5%), cottonseed
`meal (1%), corn steep liquor (0.5%), dried yeast (0.5%)
`and calcium carbonate (0.2%) at pH 6.5 was poured into
`a 500 ml-Erlenmeyer flask and sterilized at 120° C. for
`30 minutes. A loopful of slant culture of Streptomyces
`tsukubaensis No. 9993 was inoculated to the medium
`and cultured at 30° C. for four days. The resultant cul
`ture was transferred to the same preculture medium (20
`liters) in 30 liters jar~fermentor which had been steril
`ized at 120° C. for 30 minutes in advance. After the
`culture was incubated at 30° C. for 2 days, 16 liters of
`the preculture was inoculated to a fermentation medium
`(1600 liters) containing soluble starch (4.5%), corn steep
`
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`

`
`4,916,138
`
`5
`liquor (1%), dried yeast (1%), calcium carbonate
`(0.1%) and Adekanol (defoaming agent, Trade Mark,
`maker Asahi Denka Co.) (0.1%) at pH 6.8 in 2 ton tank
`which had been sterilized at 120° C. for 30 minutes in
`advance and cultured at 30° C. for 4 days.
`
`5
`
`6
`-continued
`
`Total
`
`5 g
`
`20
`
`25
`
`Isolation and Puri?cation
`The cultured broth thus obtained was ?ltered with an
`aid of diatomaseous earth (25 kg). The mycelial cake
`was extracted with acetone (500 liters), yielding 500
`liters of the extract. The acetone extract from mycelium
`and the ?ltrate (1350 liters) were combined and passed
`through a column of a non-ionic adsorption resin “Di
`aion I-IP-20” (Trade Mark, maker Mitsubishi Chemical
`Industries Ltd.) (100 liters). After washing with water
`(300 liters) and 50% aqueous acetone (300 liters), elu
`tion was carried out with 75% aqueous acetone. The
`eluate was evaporated under reduced pressure to give
`residual water (300 liters). This residue was extracted
`with ethyl acetate (20 liters) three times. The ethyl
`acetate extract was concentrated under reduced pres
`sure to give an oily residue. The oily residue was mixed
`with twice weight of acidic silica gel (special silica gel
`grade 12, maker Fuji Devison Co.), and ‘this mixture
`was slurried in ethyl acetate. After evaporating the
`solvent, the resultant dry powder was subjected to col
`umn chromatography of the same acidic silica gel (8
`liters) which was packed with n-hexane. The column
`was developed with n-hexane (30 liters), a mixture of
`30
`n-hexane and ethyl acetate (4:1 v/v, 30 liters) and ethyl
`acetate (30 liters). The fractions containing the object
`compound were collected and concentrated under re
`duced pressure to give an oily residue. The oily residue
`was mixed with twice weight of acidic silica gel and this
`mixture was slurried in ethyl acetate. After evaporating
`the solvent, the resultant dry powder was rechromato
`graphed on acidic silica gel (3.5 liters) packed with
`n-hexane. The column was developed with n-hexane
`(10 liters), a mixture of n-hexane and ethyl acetate (4:1
`v/v, 10 liters) and ethyl acetate (10 liters). Fractions
`containing the object compound were collected and
`concentrated under reduced pressure to give a yellow
`ish oil. The oily residue was dissolved in a mixture of
`n-hexane and ethyl acetate (1:1 v/v, 300 ml) and sub
`jected to column chromatography of silica gel (maker
`Merck Co., Ltd. 230-400 mesh) (2 liters) packed with
`the same solvents system. Elution was curried out with
`a mixture of n-hexane and ethyl acetate (1:1 v/v, 10
`liters and 1:2 v/ v.6 liters) and ethyl acetate (6 liters).
`Fractions containing the ?rst object compound were
`collected and concentrated under reduced pressure to
`give FR No. 900506 substance in the form of white
`powder (34 g). This white powder was dissolved in
`acetonitrile and concentrated under reduced pressure.
`This concentrate was kept at 5° C. overnight and prisms
`(22.7 g) were obtained. Recrystallization from the same
`solvent gave puri?ed FR No. 900506 substance (13.6 g)
`as colorless prisms.
`Infrared Absorption Spectrum: vmaxC-Hc13: 3680,
`3580, 3520, 2930, 2870, 2830, 1745, 1720, 1700, 1645,
`1450, 1380, 1350, 1330, 1310, 1285, 1170, 1135, 1090,
`1050, 1030, 1000, 990, 960(sh), 918 cm—1
`EXAMPLE 1
`
`45
`
`50
`
`55
`
`FR-900506 substance
`Hydroxypropyl methylcellulose 2910 (TC-5R)
`Lactose
`
`1 g
`l g
`
`65
`
`The FR No. 900506 substance (1 g) was dissolved in
`ethanol (10 ml), and thereto was added hydroxypropyl
`methylcellulose 2910 (Trade Mark: TC-SR, Maker:
`Shin-Etsu Chemical Co., Ltd.) (1 g) to prepare a suspen
`sion. To this suspension was added dichloromethane (5
`ml) to prepare a homogeneous solution. Lactose (3 g)
`was homogeneously suspended to this solution and then
`the organic solvent was removed by evaporation. The
`residual product was dried under reduced pressure for
`10 hours by vacuum dryer, milled for 2 minutes by
`coffee mill and then passed through a sieve (32 mesh) to
`give the above-identi?ed solid dispersion composition
`of the FR No. 900506 substance (5 g).
`
`EXAMPLE 2
`
`FR-900506 substance
`Hydroxypropyl methylcellulose 2910 (TC-5R)
`Lactose
`croscarmellose sodium (Ac-Di-Sol)
`
`Total
`
`I g
`l g
`2 g
`1 g
`5 g
`
`The FR No. 900506 substance (1 g) was dissolved in
`ethanol (10 ml), and thereto was added hydroxypropyl
`methylcellulose 2910 (TC-5R) (1 g) to prepare a suspen
`sion. To this suspension was added dichloromethane (5
`ml) to prepare a homogeneous solution. Lactose (2 g)
`and croscarmellose sodium (Trade Mark: Ac-Di-Sol,
`Maker: Asahi Chemical Industry Co., Ltd.) were homo
`geneously suspended to this solution and then the or
`ganic solvent was removed by evaporation. The resid
`ual product was dried under reduced pressure for 10
`hours by vacuum dryer, milled for 2 minutes by coffee
`mill and then passed through a sieve (32 mesh) to give
`the above-identi?ed solid dispersion composition of the
`FR-900506 substance (5 g).
`
`EXAMPLE 3
`
`FR-900506 substance
`Hydroxypropyl methylcellulose 2910 (TC-5R)
`Lactose
`croscarmellose sodium (Ac-Di-Sol)
`'
`
`Total
`
`1 g
`l g
`1 g
`2 g
`5 g
`
`‘The above-identi?ed solid dispersion composition of
`the FR-900506 substance‘ (5 g) was obtained in substan
`tially the same manner to that of Example 2.
`
`EXAMPLE 4
`
`FR-900506 substance
`Hydroxypropyl methylcellulose 2910 (TC-5R)
`Croscarmellose sodium (Ac-Di-Sol)
`
`Total
`
`1 g
`l g
`3 g
`5 g
`
`The FR-900506 substance (1 g) was dissolved in etha
`nol (10 ml), and thereto was added hydroxypropyl
`methylcellulose 2910 (TC-5R) (1 g) to prepare a suspen
`sion. To this suspension was added dichloromethane (5
`ml) to prepare a homogeneous solution. croscarmellose
`sodium (Ac-Di-Sol) (3 g) was homogeneously sus
`pended to this solution and then the organic solvent was
`
`NOVARTIS EXHIBIT 2037
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`

`
`4,916,138
`7
`removed by evaporation. The residual product was
`dried under reduced pressure for 10 hours by vacuum
`dryer, milled for 2 minutes by coffee mill and then
`passed through a sieve (32 mesh) to give a solid disper
`sion composition of the FR-900506 substance (5 g).
`
`8
`EXAMPLE 6
`
`FR-900506 substance
`Hydroxypropyl methylcellulose 2910 (TC-5R)
`Total
`
`1 g
`l g
`2 g
`
`The FR-900506 substance (1 g) was dissolved in etha
`nol (10 ml), and thereto was added hydroxypropyl
`methylcellulose 2910 (TC-5R) (1 g) to prepare a suspen
`sion. The organic solvent was removed by evaporation
`from the suspension, and the residue was dried under
`reduced pressure for 10 hours by vacuum dryer, milled
`for 2 minutes by coffee mill and then passed through a
`sieve (32 mesh) to give the above-identi?ed solid disper
`sion composition of the FR-900506 substance (2 g).
`What we claim is:
`1. A solid dispersion composition comprising FR
`900506 substance and water-soluble hydroxypropyl
`methylcellulose which is capable of dispersing the FR
`900506 substance, the FR-900506 substance and hydrox
`ypropyl methylcellulose being in the ratio of 1:01 to
`1:20 by weight.
`'
`2. The solid dispersion composition of claim 1, in
`which the FR-900506 substance and hydroxypropyl
`methylcellulose are in the ratio of 1:0.5 to 1:5 by weight._
`3. The solid dispersion composition of claim 2, in
`which the FR-900506 substance and hydroxypropyl
`methylcellulose are in the ratio of 1:1 by weight.
`#
`* 1k
`*
`*
`
`EXAMPLE 5
`FR-900506 substance
`Hydroxypropyl methylcellulose 2910 (TC-5R)
`The solid dispersion compositions comprising various
`ratio of the above two ingredients were obtained by the
`following method.
`
`10
`
`15
`
`20
`
`[Method]
`The FR-900506 substance (1 g) was dissolved in etha
`nol (10 ml), and thereto was added hydroxypropyl
`methylcellulose 2910 (T C-SR) (each 0.5 g, 1 g, 3 g or 5
`g) to prepare a suspension. To this suspension was
`added dichloromethane (5 ml) to prepare a homogene
`ous solution. The organic solvent was removed from
`the solution by evaporation, and the residue was dried
`under reduced pressure for 10 hours by vacuum dryer,
`milled for 2 minutes by coffee mill and passed through
`a sieve (32 mesh) to give the above-identi?ed solid dis
`persion composition in the following ratio.
`The ratio of the FR~900506 substance: hydroxypro
`pyl methylcellulose 2910 by weight are 1:05, 1:1, 1:3
`and 1:5.
`
`25
`
`30
`
`35
`
`45
`
`55
`
`60
`
`65
`
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