Development of a New Carrier for Cyclosporine A With
`Selectivity for Lymphatics
`
`K. Takada. Y. Furuya. T. Nakata. H. Yoshikawa. S. Muranishi. T. Yasumura. and T. Oka
`
`CYCLOSPORINE A (CyA) is a potent
`
`immunosuppressant with selective ac(cid:173)
`tion against T Iymphocytes. t T lymphocytes
`playa central role in the inhibition of immune
`responsiveness. and
`lymphocytes circulate
`mainly in the lymphatic systems in the body.2
`The activity of CyA is thus dependent on its
`concentration there. Based on this assump(cid:173)
`tion. we have developed a new CyA carrier
`with selective lymphatic transporting eOi(cid:173)
`cicncy. and the usefulness of this new carrier
`was studicd from both thc pharmacokinctic
`a nd pharmacologic aspects. J - S
`
`METHODS
`
`Test dosage forms arc classified into four groups: (I)
`conventional oily (CO) solution; (2) emulsion: (3) mixcd
`micellar (MM) solution; and (4) well·solubilized (W5)
`solution. Oily carrier was prepared in a 18:42:40 mixture
`of absolute elhyl alcohol. Nikkol TMGO·5 (Nikko
`Chemicals Ltd. Tokyo). and oily base. such as olive oil
`ele. Emulsion was prepared b)' dissolving CyA in oily basc
`and by dispersing into 8% (wlv) HCO-60 solution
`(polyo~yeth)'laled hydrogenated CaSler oil. Nikko Chemi·
`cals) followed by sonication at 25 °C for 5 minules. MM
`solution was prepared by dispersing linoleic acid conlain·
`ing Cyt' and HCO·60 in distilled ~'ater follo"'ed by
`sonication. W5 solution was prepared by dissolving CyA
`in 8',t, (w/v) HCO·60 or 2.0<;;' (w/v) sugar ester solution
`(OK esler. 5an·ej Chemicals Ltd. Toyonaka) follo""ed
`by sonicalion. The final CyA concentration of Ihcse test
`solutions was 3.5 mg/mL.
`The clTeet of administration route on the selective
`lymphatic delivery of CyA was studied using male Wistar
`rats in which the thoracic lymph duct ~'as cannulated
`with plastic tubing. After collcction of normal lymph.
`MM solution was administered. 7 mg/kg. by four admin·
`istration routes. intrastomach (IS). intraduodenum (ID).
`intraperitoneal (I P) and rectally. and lymph and blood
`samples were colleCTed for six hours. By centrifuging the
`blood samples at 37 ·C. plasma samples were obtained.
`CyA contenT in lymph and plasma samples was measured
`by a high· performance liquid chromatographic (l-IPLC)
`method developed here.'
`LymphaTic availabiliTY study was also performed in
`rats with the teST solutions al dose levels of 7 mg/kg. and
`the lymphatic CyA levels were measured.
`
`HeTerotopic c:Jrdiac transplantation was performed by
`intrarenal ~nastomosis of donor aorta and pulmonary
`artery to recipient aorta and inferior vena cava. respec·
`ti\·eI~·. BulTalo rats provided by Dr Ai7.awa (Hokkaido
`Uni\'ersity) were donors and Wistar rats obtained from
`5hionagi Pharmaceutical Co Ltd were recipients of heart
`allografts. Allograft recipients ~'ere given CO solution or
`8% HCO·60 solution of CyA 3t dose levels of I and 2
`mg/kg/d for I week. Rejection was defined as cessation
`of palpable heart beat and was confirmed by direct
`inspection at laparotomy and histologic examination.
`Pharmacokinetic study was performed in three groups
`of rats. One group rccciv~-d CO solution; the second group
`received 8',t, HCO·60 solution at an oral dose level of 7
`mg/kg: the third group received IV CyA dose. 3.5 mg/kg.
`Blood samples were collected for eight hours from the
`carotid artery through a cannula. and all of the gastroin(cid:173)
`testinal (Gil tract was removed at the end of the experi(cid:173)
`ment. CyA contents in plasma. GI tract. and GI contents
`~'erc measured by HPLC method . The area under the
`plasma CyA concentration \. time curve (AUC) was
`calculated by a trapezoidal rulc. All values arc repre(cid:173)
`sented as the mean =SE.
`
`RESULTS
`The rectal or IP administration of CyA in
`M M solution showed small amounts of CyA
`in the lymph for six hours. However, oral
`administration gave high lymph CyA concen(cid:173)
`trations. IS administration of CyA resulted in
`the highest CyA levels in lymph, 16 J.lgjmL.
`about twenty times higher than those obtained
`afler rectal or I P administration. The lym(cid:173)
`phatic availability of CyA was affected by its
`dosagc. As the solubili7.ation of CyA was
`accelerated. the amount of CyA transferred
`
`From /h~ D~parrmtlll of 8iopharmac~utics, Kyoto
`Pharmoc~ulical U"ivtrsity, Misosogi. Yomosillo-ku.
`Japan: and Suond Depar/ment of Surg~'y. Kyoto Pr~­
`fec/ural U"iY~rs;/y of Medicille. Hirokoji. Kamigycrku,
`Japan.
`Address rep,int requests to K. Takada. Depa,/ment of
`8iopharmaceutics, Kyoto Pharmaceutical Univtrsily.
`Misasagi. Yamasina-ku. Kyoto 607. Japan.
`6) 1987 by GrIme & Strallon, Illc.
`0041-1345/87/1901-0634$03.00/0
`
`Transplantstion ProcfltKlings. Vol XIX. No 1 (February]. 1987: pp 1711-1712
`
`1711
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`TAKADA ET Al
`
`into the thoracic duct lymph was increased.
`The rank order of lymphatic availability was
`WS solution> MM solution> emulsion>
`CO solution. The percentage of CyA trans(cid:173)
`ferred into the lymph for six hours from 8%
`H CO-60 solution was 2.14 ± 0.04%. and the
`maximum lymph CyA level was 57.10 ±
`13 .33 J.lg/mL. DK ester solution also showed
`high values. namely 1.62 ± 0.92% and
`46.35 ± 10.43 J.lg/mL. On the other hand. less
`than 0.2% of the administered CyA dose was
`transferred into the lymph from the CO solu(cid:173)
`tion.
`The immunosuppressive activity of CyA
`was increased by oral administration of CyA
`in 8% HCO-60 solution. The mean survival of
`heart allograft in rats treated with daily CyA
`doses in 8% HCO-60 solution. 2 mg/kg. for I
`week. was 21.8 ± 10.5 (SD) day. and two rat
`hearts survived more than 40 days. The mean
`survival time from the CO solution. 2 mg/
`kg/d for I week. was 12.8 ± 1.9 (SD) day.
`The control (nontreated rats) mean survival
`day was 7.6 ± 0.6 (SD) day. In the pharmaco(cid:173)
`kinetic study. the percentage recovery of CyA
`from the GI tract after oral administration of
`the CO solution was 44.5 ± 1.36%. whereas
`
`29.1 ± 3.07% was obtained after administra(cid:173)
`tion in 8% I-ICO-60 solution . The systemic
`availability of CyA. measured as the ratio.
`AUC(oral)/AUC(iv). was 28.71% for CO
`solution and 54.60% for HCO-60 solution.
`
`DISCUSSION
`HCO-60 is a well known non ionic surfac(cid:173)
`tant and is used as a pharmaceutical additive. 7
`We have been using HCO-60 as an absorption
`promoter to increase the systemic availability
`of percutaneously administered drugs.s or
`antitumor drugs9 and\to increase the lym(cid:173)
`phatic availability of interferon.lo
`In this
`experiment. both the lymphatic and systemic
`availability of CyA was increased by HCO-
`60. However, the improved lymphatic avail(cid:173)
`ability of CyA was less than 5% of the oral
`dose. and the contribution of the amount of
`CyA delivered through the lymphatic route to
`the systemic circulation was very small. On
`the other hand. the immunosuppressive activ(cid:173)
`ity of CyA was significantly increased by this
`new career. 8% HCO-60 solution. The precise
`mechanism of this intensified immunosup(cid:173)
`pressive activity of CyA is now under investi(cid:173)
`gation.
`
`REFERENCES
`
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`
`6. Takada K. Shibata N. Yoshimura H. ci al: Res
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