AU771024B2 - Compounds and compositions for delivering active agents - Google Patents

Description

Our Ref:7562890 P/00/011 Regulation 3:2

AUSTRALIA

Patents Act 1990

ORIGINAL

COMPLETE SPECIFICATION STANDARD PATENT 0 000.

000.

0 0 0 0 a a. a a Applicant(s): Address for Service: Invention Title: Emisphere Technologies, Inc 765 Old Saw Mill River Road Tarrytown New York 10591 United States of America DAVIES COLLISON CAVE Patent Trade Mark Attorneys Level 10, 10 Barrack Street SYDNEY NSW 2000 Compounds and compositions for delivering active agents The following statement is a full description of this invention, including the best method of performing it known to me:- 5020

I

p:\WPDOCS\CAB\SPECI\7562890.do-02/12/2 -1- COMPOUNDS AND COMPOSITIONS FOR DELIVERING ACTIVE AGENTS This application is a divisional of parent application No. 62756/98 (now patent No. 738735).

The ensuing description is substantially identical to the description of the specification of the parent application. The "parent" description has been readopted to facilitate identification of the parent/divisional relationship. The scope of the invention of this divisional application is set forth in the claims of this specification.

FIELD OF THE INVENTION The present invention relates to compounds for delivering active agents, and particularly biologically or chemically active agents. These compounds are used as carriers to facilitate the delivery of a cargo to a target. The carrier compounds are well suited to form non-covalent mixtures with biologically-active agents for oral administration to animals. Methods for the preparation and administration of such compositions are also disclosed.

BACKGROUND OF THE INVENTION ooooo Conventional means for delivering active agents are often severely limited by biological, chemical, and physical barriers. Typically, these barriers are imposed by the environment through which delivery occurs, the environment of the target for delivery, or the target itself. Biologically or chemically active agents are particularly vulnerable to such barriers.

For example in the delivery to animals of biologically active or chemically active pharmacological and therapeutic agents, barriers are imposed by the body.

Examples of physical barriers are the skin and various organ membranes that must be traversed before reaching a target. Chemical barriers include, but are not limited to, pH variations, lipid bi-layers, and degrading enzymes.

These barriers are of particular significance in the design of oral delivery systems. Oral delivery of many biologically or chemically active agents would be the route of choice for administration to animals if not for biological, chemical, and physical barriers such as varying pH in the gastrointestinal (GI) tract, powerful digestive enzymes, and active agent impermeable gastro-intestinal membranes. Among the numerous agents which are not typically amenable to oral administration are biologically or chemically active peptides, such as calcitonin and insulin; polysaccharides, and in particular mucopolysaccharides including, but not limited to, heparin; heparinpids; antibiotics; and other organic substances. These agents are rapidly rendered ineffective or are destroyed in the gastro-intestinal tract by acid hydrolysis, enzymes, or the like.

Earlier methods for orally administering vulnerable pharmacological agents have relied on the co-administration of adjuvants resorcinols and non-ionic surfactants such as polyoxyethylene oleyl ether and n-hexadecylpolyethylene ether) to increase artificially the permeability of the intestinal walls, as well as the co-administration of 20 enzymatic inhibitors pancreatic trypsin inhibitors, diisopropylfluorophosphate (DFF) and trasylol) to inhibit enzymatic degradation.

Liposomes have also been described as drug delivery systems for insulin and heparin. See, for example, U.S. Patent No. 4,239,754; Patel S 25 et al. (1976), FEBS Letters, Vol. 62, pg. 60; and Hashimoto et al. (1979), Endocrinology Japan, Vol. 26, pg. 337.

However, broad spectrum use of such drug delivery systems is precluded because: the systems require toxic amounts of adjuvants or inhibitors; suitable low molecular weight cargos, i.e. active agents, are not P:\WPDOCS\CAB\SPECI\7562890.dc-02/12/02 -3available; the systems exhibit poor stability and inadequate shelf life; the systems are difficult to manufacture; the systems fail to protect the active agent (cargo); the systems adversely alter the active agent; or the systems fail to allow or promote absorption of the active agent.

More recently, microspheres of artificial polymers of mixed amino acids (proteinoids) have been used to deliver pharmaceuticals. For example, U.S. Patent No.

4,925,673 describes drug-containing proteinoid microsphere carriers as well as methods for their preparation and use. These proteinoid microspheres are useful for the delivery of a number of active agents.

There is still a need in the art for simple, inexpensive delivery systems which are easily prepared and which can deliver a broad range of active agents.

SUMMARY OF THE INVENTION Compounds and compositions which are useful in the delivery of active agents are provided. These compositions include at least one active agent, preferably a biologically or chemically active agent, and compound 109 depicted below or a salt thereof.

Thus according to an aspect of the present invention as claimed there is 20 provided a composition comprising: at least one active agent; and a compound having the formula OH O N OH H 0 25 25 CI CATEMP Mmis.Div I# 0.dOc- 3/I12 -4or a salt thereof.

In one embodiment the composition of the invention may be in dosage unit form.

According to another aspect of the present invention there is provided a compound having the formula or a salt thereof.

9 9 .r Further according to another aspect of the present invention there is provided a method for preparing a composition, said method comprising mixing at least one active agent; compound 109 or a salt thereof; and optionally, a dosing vehicle.

Compounds 1-193 of the invention are represented as follows: 1 2 3 4 9@

S

S

WS.S

S.

S

5.

5565e5 S p acid -6- 0 0..

.00.

0 0*0 0 8-(3-anisoyl~aminocapryllo acid 0 .0.

4-(4-fphenoxyecetyllaminophenyl)butyic acid 4-(4-(2.nitrobonzenosulfonyl)aminophenyl)butyric acid 0 0 So: 8-(2-nitrobenzeriesutfoflyl)aminocaprytic acid OR0*0 0*10 0 6-14-(saliCYIoyl)8miflaphenlYhexaafle acid 12 13 14.

2.{(4'-salicyloyl)amilophal]cthyI methyl sulfono 16 acid 17 acid -8- OY H I 5 alicyloy3-2-gtutaryI hydrazide it 18 19 4-(4-((4-carboTYl-3-hYdrOXY PhenYI)amino)sucinYl)aminosalicyclic acid a. a. a.

a a.

N 0 0H 8-(2-pyazineborIyQlmdflocaprylic acid N00 4-t4A(2-pyrhhinecarboWlaiOphel)butvric acid 22 -9-

NO

3 0 f 6-(4-(N-2-tjitrobenzoylaminophenyl)hexanoic acid 6-14-IN-2-aminobnzoylaminphelyl)hexanoic acid 0 O A of 4-(4-(N-(6-carboxyI-2Z5-pyraziycarboxy)aminophenly)butyric acid d 4I% I 23 24- 26 4(2-Nitrobenzoylaminophe'yisuccinic acid 0 qyH

OH

OCF

3 0 8-12-(trifluoromethoxy)benzoylaoinocaprylic acid 27 10 H 28

OHOO

0 29 00 8-(benzyloxycarbOflmilO)caPYIcacid' a~i N H 0 8-(pheaoxycaboflaulifo)calPiYc acid H 31 0 320 023 0 33 II N-O

,O

4-(4-(2-hydroxynicotinoylaminaphenyl)butyric acid 1 1 OHO0 9-SalicYloylaminononarsic acid O"N0L ;QrqrOH 36

H

q henyloxycarbonylaminopheny)butyric acid 0 .37 NH. OH 3-12-msethoxybenzoylamnol)-1PropmI 00 N OH 8-(2-Hydroxynicotinoylaminocepryrc ad~dy-COOH 39.

oMe 6 -(2-vnethoxybenzoyi)amino nicotinic acid 12 0 Iao 0 salicylovlglycine N ~OH 0 0 441 42-pyrimidyl)piperazinoyl)butyric acid ao o 8-(chromone-3-carbonyl)amlnocaprylic acid N0

OH

8-(vinylbenzoyl)aminocapryic acid 00 4-4-(chramone-3-carbony)aminophenyglbutyri acid 41 42 43 44 8-cinnarnoylaminocaprylic acid 13

OH

N. H N OH 0 0 acid OH H 6OH 9-I2-hydroxybenzamidolnonandc acid 0 Ho" y 0 6-(N-4-(N-salicyloyl)aninobeazoyl*azuiocaproic acid 0

OQOH

0 4-fUavai acd 46 47 48 49 51 *fl.

I1I -cinnamnoviamlnoundecanoic acid OH H HO 0 0 4-octanoylan-o3-hydroxybenzoic acid 14- Off OHO0 I3PhenyI2.3dihydroxypropanoyl)8aminocapryrlc acid I 1 1. 52 FNt COO H 8-(N-(3-coumarincarbonyl)laminocaprylic acid Il 0 8-[N-(4-chlorobenzoytlladminocaprylic acid

S

S.

S. S *0 *6 0*

*S

S

S.

S

S.

SSWSSS

53 54 56 57 OH 0 I

OH

OH

8-(N-25-Dihydroxybenzoyl)aminocaprylic acid cto OH 8-(N-2.3-Dimethoxyb enzoyl) amino capryic acid Is 58 59 61 62 63 8 3_IN_3.5-Diacet~oxybemzoyl5II'nocaprytc acid

A.

84(N-2.4-DihydrxybezoyII6IIiocaprVe acd

CH

3 H0 00 1 Ot-(Ni-2-Methoxyanilino)3sbatic acid 16- OH H 0 6

NO

.1O-(N-2-bydroxyaniliao) sebacic acid, CH3 065

OH

2-MettioxybelZenflaiodcaloic acid 0 66 0 84(N-benzovl)WmOCaPIc acid 67 0 it 00R B-N(-loaezV)wto~r~ aci d 0 10-1-%,1%*-COOH I 69 17-

OH

HO,

8-(4-(1.2-dihydraxyethyl)benzoyl)aminocapryic acid ej IH. CO 8-[N-(4-bromobenzoyfllaminocapryic acid 71 72.

73

C

C

C

C

C

8-4N-(4odbenolOY)amilocaprytic acid 4 4 -IN-(2-iodobenzoyl)aminophenylj)butyric acid OH 0H

~H

74 -hydroxy-2-naphthoyl)aminophenyll)butyric acid -18acid 76 77 3 44-(2.4-dimethoxybenzoyl) aminophenyll propionic acid'

I

a.

oN a I4 OOH 1.

78

I

4-14-(N-4odobenzovill euninophnyl) butyric ac 00 79 344-(2.3-dimethoxybenzoyll aminophenvil orocionic acid llm OOH -4-(4-[N-(2-brombenzoyljaninopheayl) butyricad 19- 4-(4-(N-(3-bromobcnzoyl)afninophenyl]) butyric ac 81 0 Ito~ O O 0 1 Ho 8-IN-3,5 Dthydroxybmnzoyflaminocapryic acd 82 83 84 8-(N-3.5-Dimethoxy 4-hydmoxybmnzoyarinocaprylc acid

ON

OCH3 8-(N-2-6 -Dknmthoxybenzoyafracspryllc acd Br 4-(4-N-4-broobol)I~iophel)butyric acid 8-I2-hydroxy-4-chlorobenzoyllaniinacaprytic acd 86 OH 0 NH

OH

OH

8-(N-2.6-Dihydroxybenzoyl)aminocaprylic acid 87 88 8-(N-2-Hydroxy6-niethoxybenzoyl)aminocaprylic acid 0 O0 8 5 -chloro-o-anisoyl)aminocapryuic acid 09 i-go 0 OCH 3 4-(4-(Z3-imthoxbezoyI)amiophefl)butfic acid 00 OH 91 WOH3 4-(4-(5-chloro-o-anisoyl)aminophenyflbutyric acid

OH

000

N

CI

OCH

3 4 .1 4 4 -chloro..o-anisoyl)aminophenyl)butyric acid 92 -21 0 oil 8-(4-chloro-o-anisoyl)aminocaprylic acid OCR4OO 3-(4-(2.5-dimethoxybenzoy)aiophel)propioflic acid 4-{N-4-34odobenzoy)aninophenyllbuatyric acid 93 94 7 -cinnamoylaminoheptanoic acid 8-N-(34odobenzoyl)aminocapryIic acid 8.N-(4-mthoxy-3-itrobezoyI)amilOcapryic acid 96 97 98 22 99 B-N-(Z-'mthoxy- 4 -nitrobenzoyllaminocaprylic acid OMe

N.

4 4 2 5 -dimethoxybenzoyl)aminopheny)butyic acid 100 101

OH

Br 102

I-

2 -hydroxy-5-bromobenzoyg)aminocaprylic ac 103 104 3 -Indolebutyric acid 23 Y 0H 105 cr0O 106 gNH NHi OCHY 107 4-[4-{2,6-dmethoxybcnoy)aminophnylJ butyric acid .108 *100 4~~-[N-4mylhoxy--ib'obemoylmnopeui add -24- 0 N H O 1 12 113 8-(N2-hdroX-4-itrbenzyl~minoaprticacid *0 9 to0 0* 9.

vP acid 114 115 0" 8-(rJ-3-methylsalicyloyl)aminocapryrnc acid 25 N OH 116 8 -(N-5-methylsalicyloyl)aminocaprylic acid 0 117 9-(cinnamoylamino)nonanoic acid 0 OH 11-8 4-(4-(2-chloro-5-nitrobenzoyliaminophenyl)butyric acid of( off 119 4-(-[N-(2-hydroxy-5-iodobenzoylflaminophanyllbutyrc acid 0 120 2 H H N-2-nitiophenyl-N'-(8-octanoic acid) urea H 121 sebecoyl amide acid -26-- I 122 -8-[N-(2-hyr-3dicorobzoyl)jaminocapylic ai .123 F' cow

HH

8 N-(2-chydro-3-flroby~aminocapryc acid ol 126

COOH

8--2-drox-3-nrobnz~yincapryic acid 112 4-(4-Salicyloylaninophenyfl-4-oxobutyric acid -27- 128 l 2 -cmnnanoyldodecanoic acd H H.129 0 4 -1 4 -4N-(3-hYdoxY-2-naPhthoYl)amuanophenylj Ibutyric acid 0 130 0 0 8 44-chloro-3-nitirobenzoyllaminocapgyllc acd 0 131 4-chldoroncotkuenoywWiocapryc acid 0 I 133 %Th{iJ 0

CO

2

H

4-(4-phtharamidophenyl)butyric acd -28- 134 acid 135 lyl)propioflic acd 136

N-,--COOH

II

137- 138 139 8-(2-Il .2-dlhydralsolndole-1 -one))octanolc acid IHO0

OH

8-(N-1 hydroxy-2-nphthoylaiocPYic acid -29- 0 00~ 8.(phthalimido)capryic acid 140 141 1 O-I4-chtoro2-hydroxyaflilo)sebacic acid monoamide o* 0 0 0 142 143 144 4-(4-(4.choro-3-nitrobelzoyi)milophel)butyric acid OH 0 0 I1-N-(1 -hydroxy-2-naphthoy)aminouldcaloic acid 1,Bis(N-2crboxylphenyl-N-IW*-8octanoic aidluiresI~oxalyl diamide 145 146 147 2-(2I'l-t2-chorobenzoyluaiinothoxylthanalI 2-(2-N-(4-ch~orobenzoylaminothoxyI ethanol I. I I I I. I I I

I.

I

I

I. I I I

II

I

4-(2-Jicthoxybenzoylamino)phenyl 2-caztxyediyl su-Ifoxide 0 0 0% 4-(2-.nwthoxybmwnzyaino)phenyi 2-carboxycthyl sulfone OH 0 0NQ 0 C0 2 H1 4-(4-(3-hydroxyphthhtimido)phenVI)butynic acid 148 149 150 -31 24-2-N-(2-methoxybnzoy)minethXY~ethaiIol

H

151 152 153 4* 4* 4. 4 4* 4* 4**4 4 4 4* 4 4* 4.

4***44 4 Bis(N-2-carboxyplienyt-N-INW-3(4-aminoph-enyllpropionic acidiurealloxaylyl diamide 0 c o zti1 5 4 trans4-(2-anunobcnzamnidomthyl)cyclohexaaocaboxyli: acid 11 -N-(3.5-dichloro-2.hydroxybenzoyflaminoundecanfle acd 155 32

~~OH

156 157 2-(N-(2-bromobenzoyl)aminoetkioxylethanoI 0 0 C

OH

OH

CI

7-dichloro-2-byro2-yenoxybe4(4aminohetnoluic acid 158.

.Of S 00* :*toot 06 159 160 carboxylic acid N-13.5-dichloro-2-l1ydroxybelzoy3(4-milophel)propioflic acid IC L A- J- H l 161 12 -35dclr--vrxyezy~mnddcni acid -33- OH H 1 0 N-(2-hydroxy-4-carbxyphcnyl)-6-Iarnamidd N-2-browbenzoylmwophorme

HO

162 163 164 165 166 0 2-'-24odobenzoyl)aninoelhoxvjeihanol OH H 0

OH

5-(4-ctdoro-2-hydroxyanlinocarbonyllvaleric acid 0

OH

8-(2-hydroxyphelcty)-am2foraprYlic acid 167 34 Q N4 CO 2

H

OH

N-Saficoyl-5-(3-aminophenyl)vateric acid 0

OH

4-(4-(2-cdioxylbenmy1aminopheuyI)butyric acid OH H 0

OH

9-2(-yrxlyiyaiaabnl nonaniC acid 168 169 170-

A,

171 172 .2-IN-(2-hydroxybenzoylaminlOetoxylethaloI 35 0OH 0 173 cl-I I 4-[N -3.5-drichloro-2-hydroxybenzoyIJlamnaphenylacetic acid H It H174 8-12-hdroxy5chloamli~gnocabomylUoctalol acd 0 COIH 175 N.,saricoyI5-4aminopheny)valeric acid 114 0*176 S-12-hydmoxy-5-mothylalhlinocarboflylnofafloic acid I 177 5-t2-hydroxy-5-rnethyaliocrboyl)vaIeic acid 36 178 179 180 181 3-f3-(salicyioyflamlophenyflpropionic acd 0 J OH .ocRzi- 0

S

S.

S

S.

S

8-(2-.thoxybenzoyllwminocaprylic acid

NH

8-(3-Phmnoxylpropionylarino)caprylic acid 182 37 4-(Saicy[OVI)asminophenylethyltebrazola 183 184 185 186

S-

4 -(4-N-salicyloyl)anminophcnyl)butyric) amiinocaprylic acid

H

F J Q. 0 OH 0 H 4-(4-N-(4--4N-(2-Fluorocinnamyl)aminophenyl)butyric)amfliophefl)butyric acid a a. a a.

a.

a a a 4-INSNSuricyloyt)anminocapryic)8mtnopheny)butyrlc acid 187 38 188 189 0 ~WO acd 0 8 -Sn 4 .thylsalicyloyl)aminocapryljc acid 190 191 192

S

10-N(2-ydroy-5nitrmilno)-o-oode..iacid 193 4 4 2 -chloronicotinoyllaminoplenybutyri acid -39- Compositions comprising the carrier compounds discussed above and active agents are effective in delivering active agents to selected biological systems.

Thus according to yet another aspect of the present invention as claimed there is provided a method for administering a biologically-active agent to an animal in need of said agent, said method comprising administering orally, intranasally, sublingually, intraduodenally, subcutaneously, rectally, vaginally, bucally or ophthalmically to said animal a composition of the invention.

According to another aspect of the present invention there is provided a method for passing a biologically active agent across the blood/brain barrier of an animal in need of said agent, said method comprising administering to said animal a composition of the invention.

Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

DETAILED DESCRIPTION OF THE INVENTION The specific compositions of the present invention include an active agent and a carrier. These compositions may be used to deliver various active agents through S various biological, chemical, and physical barriers and are particularly suited for delivering active agents which are subject to environmental degradation. The compositions of the subject invention are particularly useful for delivering or administering biologically or chemically active agents to any animals such as birds including, but not limited to, chickens; mammals, such as primates and particularly humans; and insects.

•.Other advantages of the present invention include the use of easy to prepare, inexpensive raw materials. The compositions and the formulation methods of the present invention are cost effective, simple to perform, and amenable to industrial scale up for commercial production.

Subcutaneous, sublingual, and intranasal coadministration of an active agent, such as, for example, recombinant human growth hormone (rhGH); salmon calcitonin; heparin, including, but not limited to, low molecular weight heparin; parathyroid hormone; and compounds in compositions as described herein result in an increased bioavailability of the active agent compared to administration of the active agent alone.

Active Agents Active agents suitable for use in the present invention include biologically or chemically active agents, chemically active agents, including, but not limited to, fragrances, as well as other active agents such as, for example, cosmetics.

Biologically or chemically active agents include, but are not limited to, pesticides, pharmacological agents, and therapeutic agents. For example, biologically or chemically active agents suitable for use in the present invention include, but are not limited to, peptides, and particularly small peptides; hormones, and particularly hormones which by themselves do not or only a fraction of the administered dose passes through the gastro-intestinal mucosa and/or are susceptible to chemical cleavage by acids and enzymes in the gastro-intestinal tract; polysaccharides, and particularly mixtures of mucopolysaccharides; carbohydrates; lipids; or any combination thereof. Further examples include, but are not limited to, human growth hormones; bovine growth hormones; growth t releasing hormones; interferons; interleukin-1; interleukin II; insulin; heparin, and particularly low molecular weight heparin; calcitonin; erythropoietin; atrial naturetic factor; antigens; monoclonal antibodies; somatostatin; adrenocorticotropin, gonadotropin releasing hormone; oxytocin; vasopressin; cromolyn sodium (sodium or disoditim chromoglycate); vancomycin; desferrioxamine (DFO); parathyroid hormone; antimicrobials, including, but not limited to anti-fungal agents; or any combination thereof.

Particular mention is made of calcitonin as the active agent.

C.

Carriers Although compounds 1-193 above have been found to act as carriers for the oral delivery of biologically or chemically active agents, special mention is made of compounds 9, 35, 64, 67, 79, 102, 109, 111, 117, 122, 136, and 141, above, and in particular compound 109.

-41 *b Properties of compounds 1-193 are listed in Table 1, below.

TABLE 1 Carrier Properties Melting Compound Anal. Calculated For Found Point oCi CH N S C H N S 1 48.8 4.70 4.40 48.81 4.64 4.39 2 64.73 7.97 10.06 64.54 7.81 10.19 3 55.33 5.80 4.03 55.40 5.79 3.96 69-71 4 62.64 6.06 5.62 62.75 6.08 5.51 151-154 65.16 6.11 13.40 65.29 6.03 13.29 144-145 6 54.70 3.24 3.75 54.29 3.24 3.54 165-169 7 69.00 6.11 4.47 69.09 6.24 4.43 126-129 8 65.51 7.90 4.78 65.60 8.25 4.83 89-90 9 68.99 6.11 4.47 69.01 6.08 4.47 104-107 52.74 4.42 7.69 52.91 4.45 7.49. 142-145 11 48.83 5.85 8.14 48.95 5.89 8.02 120-122 12 69.71 6.47 4.28 69.56 6.47 4.38 144-146 13 65.51 7.90 4.77 65.23 7.88 4.72 72.5- 74.5 14 60.17 5.36 4.39 10.04 60.09 5.36 4.35 9.99 155-156 52.38 4.79 11.11 52.45 4.94 11.08 220-222 16 67.60 5.95 3.94 67.34 6.01 3.91 219-222 17 68.09 6.53 3.78 67.77 6.24 3.81 130-133 18 54.13 5.30 10.52 54.12 5.24 10.54 192.5- 195.5 19 55.26 4.21 7.16 54.48 4.32 6.86 >280 dec 65.51 7.90 4.77 65.52 7.90 4.77 75-80 21 58.85 7.21 15.84 58.86 7.16 15.69 120-122 22 63.15 5.30 14.73 63.30 5.43 14.18 197-201 -42- TABLE 1 Carrier Properties Melting Compound Anal. Calculated For Found Point C H N S C H N S 23 64.04 5.66 7.86 64.17 5.67 7.75 188-190 24 69.91 6.88 8.46 69.98 6.79 8.58 131-134 58.36 4.56 12.76 58.20 4.63 12.61 138-141 26 56.98 3.94 7.82 56.39 3.92 7.74 221-223 27 55.33 5.80 4.03 55.47 6.10 4.04 70-72 28__ 29 65.74 7.58 4.79 65.51 7.89 4.78 52-55 64.50 7.57 5.02 64.07 7.81 5.40 70-74 31 54.70 5.17 3.99 54.50 4.99 3.95 173-174 32 58.63 5.94 9.12 58.73 6.20 10.34 125-129 33 69.00 6.10 4.47 69.18 6.08 4.54 100-102 34 63.99 5.37 9.33 63.46 5.35 9.06 218- 221 c 35 65.5 7.90 4.78 65.37 8.00 4.66 96-97C 36 68.22 5.72 4.68 67.88 5.65 4.55 134-137 37 63.14 7.23 6.69 63.15 7.29 6.58 53.5-56 38 60.00 7.14 10.00 59.78 7.31 9.94 135-138 39 61.67 4.41 10.29 61.69 4.41 10.12 >225 40 55.39 4.65 7.18 55.52 4.77 7.30 162.5- 166 41 56.10 6.52 20.14 55.66 6.71 19.69 129-131 42 65.24 6.39 4.23 65.42 6.16 3.78 130- 133.5 43 70.59 7.96 4.84 70.35 8.13 4.79 111-113 44 68.37 4.88 3.99 68.61 4.89 3.79 120-123 70.59 7.96 4.84 70.48 7.97 4.71 108-110 46 60.75 6.37 5.90 60.97 6.18 5.80 100.5- 103 -43- TABLE 1 Carrier Properties Melting Compound Anal. Calculated For Found Point c C H N S C H N S 47 64.50 7.57 5.02 64.42 7.58 5.01 97-100 48 64.86 5.98. 7.56 64.50 6.01 7.52 165-169 49 72.18 3.76 0.00 72.13 3.84 0.00 >225 72.51 8.76 4.23 72.39 8.84 4.12 120-122 51 64.50 7.58 5.01 64.75 7.65 4.69 200.5- 204 52 7.74 4.33 7.82 4.30 88-89 53 65.24 6.39 4.23 65.15 6.46 4.23 93-97 54 60.49 6.77 4.70 60.54 6.76 4.65 114-116 6404 7.17 4.98 63.90 7.11 4.93 105-106 56 61.00 7.17 4.74, 60.49 6.92 4.65 146-148 57 63.14 7.79 4.33 63.22 7.82 4.36 59-61 58 63.14 7.79 4.33 63.17 7.86 4.26 102-104 59 63.14 7.79 4.33 63.35 7.68 4.20 89-90 60 60.15 6.64 3.69 59.84 6.66 3.64 112-113 61 65.53 8.85 6.65 65.34 8.73 6.67 89-92 62 61.00 7.17 4.74 60.94 7.12 4.49 104-108 63 66.43 8.20 4.56 66.29 8.23 4.36 77-78 64 65.51 7.90 4.77 65.52 8.06 4.54 97-98 69.59 9.28 4.77 69.64 9.35 4.86 62-65 66 68.41 8.04 5.32 68.41 8.06 5.28 88-89 67 62.12 7.49 4.53 61.94 7.45 4.43 98-99 68 64.04 7.17 4.98 64.07 7.16 4.95 106-107 69 52.64 5.89 4.09 52.63 5.85 4.03 109-110 63.15 7.74 4.33 63.26 7.90 4.14 97-100 71 52.64 5.89 4.09 52.67 5.99 3.97 114-115 -44- TABLE 1 Carrier Properties Melting Compound Anal. Calculated For Found Point C H N S C H N S 72 46.31 5.18 3.61 46.25 4.86 3.52 143-144 73 49.89 3.94 3.42 49.92 3.85 3.39 170-171 74 72.19 5.48 4.01 71.51 5.33 3.75 180 66.46 6.16 4.08 66.47 6.26 4.06 168.5- 171 76 67.37 5.26 4.91 67.31 5.25 5.07 130-133 77 65.65 5.78 4.26 65.49 6.04 4.26 179-183 78 49.89 3.94 3.42 49.8 3.71 3.29 237-238 79 65.65 5.78 4.26 65.21 6.05 4.24 156-158 56.38 4.45 3.87 56.4 4.21 3.91 130-131 81 56.38 4.45 3.87 56.46 4.5 3.84 197-198 82 56.6 7.49 4.4 56.3 7.49 4.14 58-62 83 57.03 8.2 3.91 57.17 7.8 3.7 138-140 84 57.58 7.11 3.95 57.52 7.7 3.94 85 56.38 4.45 3.87 56.31 4.25 3.64 230-231 86 57.42 6.42 4.46 57.14 6.45 4.2 116-117 87 61 7.17 4.74 61.18 7.05 4.65 108-109 88 62.12 7.49 4.53 62.34 7.21 4.39 107-109 89 58.63 6.76 4.27 58.53 6.81 4.2 117-118 66.46 6.16 4.08 66.18 6.15 3.84 100-104 91 62.16 5.21 4.03 61.93 4.97 3.86 183-185 92 62.16 5.21 4.03 62.2 5.14 3.98 167-170 93 58.63 6.76 4.27 58.64 6.83 4.19 106-108 94 65.65 5.81 4.25 65.56 5.64 4.2 153-156 49.89 3.94 3.42 49.9 3.81 3.18 216-217 96 69.82 7.64 5.09 69.91 7.66 5.02 129-131 *6 *r TABLE 1 Carrier Properties Melting Compound Anal. Calculated For Found Point (OC) C H N S C H N S 97 46.31 5.18 3.61 46.54 4.95 3.64 122-123 98 56.8 6.55 8.28 56.69 6.67 8.1 99 56.8 6.55 8.28 57.37 6.57 8.33 117-118 100 60.33 5.06 7.82 59.98 4.97 7.67 207-209 101 66.46 6.16 4.08 66.37 6.32 3.96 126-128 102 50.29 5.63 3.91 50.14 5.7 3.76 129-131 103 70.93 5.95 6.89 70.94 6.44 6.89 104 65.84 6.14 8.53 65.94 6.19 8.54 228-231 105 64.96 5.77 8.91 64.89 5.82 8.82 106 66.65 6.48 8.18 66.39 6.49 8.05 140-142 107 66.47 6.12 4.07 66.5 6.26 4.08 140-142 108 60.33 5.06 7.82 60.32 4.99 7.78 150-151 109 57.41 6.42 4.46 57.07 6.44 4.39 121-123 110 44.46 4.97 3.46 133-135 111 69.28 7.03 4.25 68.86 7.07 4.11 147-149 112 55.55 6.22 8.64 55.27 5.99 8.5 120-121 113 53.99 4.26 3.7 53.98 4.25 3.63 210 decom 114 57.49 7.39 4.74 57.72 7.57 4.43 80-83 115 65.5 7.9 4.77 64.97 7.79 4.75 90-92 116 65.5 7.9 4.77 65.11 8.03 4.71 125-127 117 71.26 8.3 4.2 70.6 7.89 4.83 94-96 118 56.29 4.17 7.72 56.23 4.01 7.6 173-175 119 47.89 3.81 3.29 47.52 3.71 3.16 236-237 120 55.7 6.55 13 55.71 6.58 13.05 123-5 121 57.98 5.81 7.95 57.9 7.11 7.82 131-133

S

9 9 000S

S

9 0C.

.i

S.

9

S

es..

0 5 -46- TABLE 1 Carrier Properties Melting Compound Anal. Calculated For Found Point (0c) C H N S C H N S 122 51.74 5.5 4.02 51.41 5.43 3.61 118- 119.5 123 41.22 4.38 3.2 41.45 4.36 2.94 143- 144.5 124 57.06 6.06 4.44 57.02 6.12 4.35 57-58 125 61.18 4.83 4.2 60.71 4.76 3.89 214 decom 126 55.55 6.22 8.64 55.4 6.24 8.53 150-151 127 65.17 4.83 4.47 65.27 4.87 4.48 208-209 128 73.03 8.99 4.06 72.92 9.36 4.1 99-101 129- 72.25 5.44 4 72.14 5.24 4.01 216-217 130 52.56 5.58 8.17 52.66 5.44 8.21 96-100 131 56.28 6.41 9.38 56.32 6.42 9.28 98-100 132 52.56 5.58 8.17 52.46 5.65 7.86 150-153 133 69.89 4.89 4.53 69.64 5 4.54 136-9 134 71.68 5.2 4.2 71.24 5.1 4.13 251-253 135 65.64 5.78 4.25 65.3 5.91 4.04 79-83 136 33.92 3.61 2.64 34.48 3.84 2.48 164-165 137 57.06 6.06 4.44 57.09 6.17 4.45 88-89 138 69.79 7.69 5.09 69.68 7.78 5.08 102-3 139 69.28 7.04 4.25 68.99 7 4.1 107-108 140 66.42 6.62 4.84 66.2 6.49 4.81 88-9 141 58.62 6.76 4.27 58.66 6.93 4.18 134-135 142 63.38 7.21 5.28 63.22 7.28 5.24 71-73 143 56.29 4.17 7.72 56.19 4.04 7.65 156-160 144 71.13 7.88 3.77 70.39 7.91 3.64 95-97 145 58.44 6.06 8.02 58.25 6.38 7.84 165-8 *a a -47 TABLE 1 Carrier Properties Melting Compound Anal. Calculated For Found Point 0

C)

C H N S C H N S 146 54.22 5.79 5.75 54.26 5.65 5.69 77-78.5 147 54.22 5.79 5.75 54.21 5.85 5.61 80-81 148 58.78 4.93 40.3 58.64 4.89 3.97 172-173 149 56.19 4.72 3.85 56.31 4.67 3.86 177 150 66.46 4.65 4.31 66.41 4.56 4.23 158-160 151 58.61 7.24 5.69 58.79 7.35 5.66_ 152 54.22 5.79 5.75 54.21 5.72 5.62 54-55 153 60.85 4.25 7.89 60.27 4.37 7.89 >260 154 62.5 7.3 10.14 64.77 7.27 9.9 187-190 155 55.4 6.5 3.6 55.56 6.51 3.5 114-116 156 45.85 4.9 4.86 46.06 4.78 4.71 67-68 156 48.8 4.7 4.4 48.81 4.64 4.39 144-146 157 50.3 5.1 4.2 50.25 5.12 3.99 141-143 158 55.5 4.1 3.8 55.55 3.88 3.75 190-192 159 64.97 6.9 5.05 64.7 6.82 5.02 171-174 160 54.3 3.7 4 54.31 3.58 3.83 222-224 161 56.4 6.7 3.5 56.69 6.98 3.11 76-78 162 63.63 6.47 5.3 64.76 6.84 4.74 188-191 163 48.91 4.48 5.19 48.89 4.31 5.10 88.5-90 164 66.66 10.0 5.18 66.69 10.7 5.16 67.5- 4 7 70.5 165 39.42 4.21 4.18 39.19 4.35 3.88 oil 166 53.05 5.19 5.16 53.06 5.03 4.86 151-152 167 65.53 7.85 4.78 65.4 7.84 4.57 85-89 168 68.99 6.11 4.47 68.62 5.87 4.49 162-6 169 69.71 6.47 4.28 69.67 6.58 4.50 132.5- 135 -48- TABLE 1 Carrier Properties Melting Compound Anal. Calculated For Found Point (c) C H N S C H N S 170 61.21 7.53 9.52 61.21 7.68 9.46 134-135 171 62.14 7.44 4.53 61.96 7.52 4.57 101-104 172 58.63 6.71 6.22 58.15 6.83 6.04 173 52.96 3.26 4.12 52.96 3.28 4.02 225-227 174 57.42 6.42 4.46 57.3 6.38 4.39 119-120 175 68.99 6.11 4.47 68.84 6.08 4.51 131-4 176 66.43 8.2' 4.56. 66.42 8.16 4.51 109-110 177 62414i 6.82 5.57 61.96 6.66 5.52 127-128 178 51.00 4.56 3.97" 51.09 4.61 3.93 179 67.36 5.30 4.90 67.26 5.24 4.91 185-186 180 66.43 8.20 4.56 66.32 8.60 5.12 51.5-55 181 69.92 6.79 8.58 67.02 6.93 8.20 81-84 182 66.46 8.14 4.56 66.43 8.34 4.47 82-84 183 62.13 4.89 22.64 62.05 4.88 22.45 271-272 184 68.16 7.32 6.36 67.73 7.44 6.70 114-117 185 71.30 5.98 5.73 71.10 5.97 5.74 146-149 186 68.16 7.32 6.36 67.94 7.31 6.41 105-108 187 65.51 7.90 4.77 65.35 7.63 4.59 102-103 188 64.50 7.58 5.01 64.19 7.69 4.83 133-134 189 64.5 7.58 5.01 64.5 7.57 4.90 116-118 190 61.15 7.71 3.97 61.27 7.79 4.08 124-127 191 65.5 7.9 4.77 65.32 7.94 4.7 114-115 192 56.77 6.51 8.28 56.83 6.76 8.21 141-143 193 60.29 4.74 8.79 60.17 4.58 8.74 202-205 194 48.8 4.7 4.4 48.81 4 4.39 144-146 1 .64 -49- These carrier compounds or poly amino acids, and peptides, including the amino acids, may be used to deliver active agents including, but not limited to, biologically or chemically active agents such as for example, pharmacological and therapeutic agents.

An amino acid is any carboxylic acid having at least one free amine group and includes naturally occurring and synthetic amino acids.

Poly amino acids are either peptides or two or more amino acids linked by a bond formed by other groups which can be linked, e.g. an ester, anhydride, or an anhydride linkage.

Peptides are two or more amino acids joined by a peptide bond.

Peptides can vary in length from dipeptides with two amino acids to poly peptides with several hundred amino acids. See Chambers Biological Dictionary, editor Peter M. B. Walker, Cambridge, England: Chambers Cambridge, 1989, page 215. Special mention is made of di-peptides, tri-peptides, tetra-peptides, and penta-peptides.

Salts such as, for example, sodium salt of these carrier compounds can be used as well.

Many of the compounds described herein are derived from amino acids.

20 Many of the compounds of the present invention can be readily prepared from amino acids including, but not limited to, aminocaprylic acid, butyrylhydroxaminic acid, aminophenylbutyric acid, aminophenylhexanoic acid, aminophenylpropionic acid, amino salicylic acid, aminophenylsuccinic acid, aminononanic acid, aminonicotinic acid, amino valenic acid, 25 aminophenylacetic acid, aminocaproic acid, aminoundecanoic acid, aminoheptanoic acid, aminohydroxybenzoic acid, and aminodecanoic acid by methods within the skill of those in the art based upon the present disclosure and the methods described in U.S. patent application serial nos. 60/017,902, filed March 29, 1996; 08/414,654, filed March 31, 1995; 08/335,148, filed October 25, 1994; and 60/003,111, filed September 1, 1995.

For example, these compounds may be prepared by reacting the single acid with the appropriate agent which reacts with free amino moiety present in the amino acids to form amides. Protecting groups may be used to avoid unwanted side reactions as would be known to those skilled in the art.

The carrier compound may be purified by recrystallization or by fractionation on solid column supports. Suitable recrystallization solvent systems include acetonitrile, methanol and tetrahydrofuran. Fractionation may be performed on a suitable solid column supports such as alumina, using methangl/n-propanol mixtures as the mobile phase; reverse phase column supports using trifluoroacetic acid/acetonitrile mixtures as the mobile phase; and ion exchange chromatography using water as the mobile phase. When anion exchange chromatography is performed, preferably a subsequent 0-500 mM sodium chloride gradient is employed.

Delivery Systems The compositions of the present invention may include one or 20 more active agents.

In one embodiment, compounds or salts of compounds 1-193 or poly amino acids or peptides that include at least one of these compounds or salts may be used directly as a delivery carrier by simply mixing one or more compound or salt, poly amino acid or peptide with the active agent prior to 25 administration.

The administration mixtures are prepared by mixing an aqueous solution of the carrier with an aqueous solution of the active ingredient, just prior to administration. Alternatively, the carrier and the biologically or chemically active ingredient can be admixed during the manufacturing 51:- process. The solutions may optionally contain additives such as phosphate buffer salts, citric acid, acetic acid, gelatin, and gum acacia.

Stabilizing additives may be incorporated into the carrier solution.

With some drugs, the presence of such additives promotes the stability and dispersibility of the agent in solution.

The stabilizing additives may be employed at a concentration ranging between about 0.1 and 5 preferably about 0.5 Suitable, but non-limiting, examples of stabilizing additives include gum acacia, gelatin, methyl cellulose, polyethylene glycol, carboxylic acids and salts thereof, and polylysine. The preferred stabilizing additives are gum acacia, gelatin and methyl cellulose.

The amount of active agent is an amount effective to accomplish the purpose of the particular active agent. The amount in the composition typically is a pharmacologically, biologically, therapeutically, or chemically effective amount. However, the amount can be less than a pharmacologically, biologically, therapeutically, or chemically effective amount when the composition is used in a dosage unit form, such as a capsule, a tablet or a liquid, because the dosage unit form may contain a multiplicity of carrier/biologically or chemically active agent compositions or 20 may contain a divided pharmacologically, biologically, therapeutically, or chemically effective amount. The total effective amounts can then be administered in cumulative units containing, in total, pharmacologically, e- biologically, therapeutically or chemically active amounts of biologically or pharmacologically active agent.

S 25 The total amount of active agent, and particularly biologically or chemically active agent, to be used can be determined by those skilled in the art. However, it has surprisingly been found that with some biologically or chemically active agents, the use of the presently disclosed carriers provides extremely efficient delivery, particularly in oral, intranasal, sublingual, 52 intraduodenal, rectal, vaginal, buccal, ophthalmic, or subcutaneous systems as well as systems for crossing the blood/brain barrier. Therefore, lower amounts of biologically or chemically active agent than those used in prior dosage unit forms or delivery systems can be administered to the subject, while still achieving the same blood levels and therapeutic effects.

The amount of carrier in the present composition is a delivery effective amount and can be determined for any particular carrier or biologically or chemically active agent by methods known to those skilled in the art.

Dosage unit forms can also include any of excipients; diluents; disintegrants; lubricants; plasticizers; colorants; and dosing vehicles, including, but not limited to water, 1,2-propane diol, ethanol, olive oil, or any combination thereof.

Administration of the present compositions or dosage unit forms preferably is oral or by intraduodenal injection.

The delivery compositions of the present invention may also include one or more enzyme inhibitors. Such enzyme inhibitors include, but are not limited to, compounds such as actinonin or epiactinonin and derivatives thereof. These compounds have the formulas below: S H NHOH 2

HO

Me Actinonin 5.3- NH OH

HO

Me Epiactionin Derivatives of these comnpounds are disclosed in U.S. Patent No. 5,206,384.

Actinonin derivatives have the formula: 0

C,

PH

2 H 0 0 CH *CH(

CH

3 R6 wherein R' is sulfoxymethyl -or carboxyl or a substituted carboxy group 25 selected from carboxamide, hydroxyaminocarbonyl and alkoxycarbonyl groups; and R' is hydroxyl, alkoxy, hydroxyamino or sulfoxyamino group.

Other enzyme inhibitors include, but are not limited to, aprotinin (Trasylol) and Bowman-Birk inhibitor.

-54- The compounds and compositions of the subject invention are useful for administering biologically or chemically active agents to any animals such as birds; mammals, such as primates and particularly humans; and insects. The system is particularly advantageous for delivering chemically or biologically or chemically active agents which would otherwise be destroyed or rendered less effective by conditions encountered before the active agent its target zone the area in which the active agent of the delivery composition are to be released) and within the body of the animal to which they are administered. Particularly, the compounds and compositions of the present invention are useful in orally administering active agents, especially those which are not ordinarily orally deliverable.

DESCRIPTION OF THE PREFERRED EMBODIMENTS The following examples illustrate the invention without limitation.

All parts are given by weight unless otherwise indicated.

Example 1 Carrier Preparation General Preparations of Carriers. The following procedures were used to prepare the compounds described herein. Many of the compounds 20 were prepared by reaction of the appropriate amino acid with the appropriate acid chloride. The preparation of compound 79 is given as a representative example of the compounds prepared in this manner.

99 Preparation of Compound 79. Method A. A 1 L round bottom 25 flask fitted with a magnetic stirrer was charged with 3-(4aminophenyl)propionic acid (46.3 g, 0.28 moles, 1.17 equiv.) and 2 M aqueous sodium hydroxide (300 mL). 2,3-dimethoxybenzoylchloride (48.0 g, 0.24 moles, 1.00 equiv.) was added portionwise over 1 h to the stirred solution. After the addition, the reaction was stirred for 2.5 h at ambient temperature, and the pH of the solution was kept at ca 10 by the addition of M sodium hydroxide. The solution was then acidified with 1 M hydrochloric acid (3 x 100 mL), water (100 mL), and air dried. It was redissolved in boiling acetone (ca 500 mL), decolorized with activated charcoal and filtered. Water (1.5 L) was added to the filtrate to induce the formation of a brown oil. The brown oil solidified upon stirring at room temperature for 10 min. The crude solid was collected by filtration and recrystallized from 70% methanol-water to afford compound 79 as a tan solid (39.5) g, Compounds 1, 5, 30, 31,33, 36, 53-66, 68, 69, 71-74, 78, 88, 95, 97-99, 102, 108-110, 112-115, 119, 121-126, 136, 137, 139, 141, 144, 146, 147, 151, 152, 155-158, 160, 161, 163, 165, 166, 170, 172- 174, 176, 177, 184-186, 188, 189, 191 and 192 were also prepared by this process.

Preparation of Compound 79. Method B. A 2 L three-neck round bottom flask was fitted with a magnetic stirrer and two addition funnels under an argon atmosphere. A suspension of 3-(4-aminophenyl)propionic acid (46.3 g, 0.28 moles, 1.17 equiv.) in ethyl acetate (700 mL) was added to the 20 flask. A solution of 2,3-dimethoxybenzoylchloride (48.0 g, 0.24 moles, 1.00 equiv.) in ethyl acetate (250 mL) was charged to one of the addition funnels and added dropwise over 1 h. Triethylamine (28.20 g, 0.28 moles, 1.00 0*00 ,equiv.) was subsequently charged to the second funnel and added dropwise over 15 min. The reaction was stirred at ambient temperature for 3 and s 25 the solvent was evaporated in vacuo giving a residual brown oil. Water (600 mL) was added to the residue followed by sodium hydroxide (2 M, 500 mL), and the mixture was stirred at ambient temperature for 3 hours. The resultant brown solution was acidified with 2 M hydrochloric acid (ca 1 L).

After cooling the mixture in an ice bath for 1 h, a yellow solid formed and -56was collected by filtration. The solid was washed with water (3 x 1.5 L) and recrystallized from 50% ethanol-water to give compound 79 as a tan solid (59.2 g, 68%).

Compounds 18, 32, 37, 41, 168, 175, and 183 were also prepared by this process.

Preparation of Compound 79. Method C. A 2 L round bottom flask equipped with a magnetic stirrer and a reflux condenser was charged with a suspension of 3-(4-aminophenyl)propionic acid (46.3 g, 0.28 moles, 1.17 equiv.) in dichloromethane (560 mL). Chlorotrimethylsilane (62.36 g, 0.57 moles, 2.05 equiv.) was added in one portion, and the mixture was heated to reflux for 1 h under argon. The reaction was allowed to cool to room temperature and was placed in an ice bath (internal temperature 0 The reflux condenser was replaced with an addition funnel 15 containing triethylamine (42.50 g, 0.42 moles, 1.50 equiv.). The triethylamine was added dropwise over 15 min, and a yellow solid formed during the addition. The funnel was replaced by another addition funnel containing a solution of 2,3-dimethoxybenzoylchloride (48.0 g, 0.24 moles, 1.00 equiv. in dichloromethane (100 mL). The solution was added dropwise 20 over 30 min. The reaction was stirred in the ice bath for another 30 min and at ambient temperature for 1 h. The dichloromethane was evaporated in vacuo to give a brown oil. The brown oil was cooled in an ice bath, and an ice-cold solution of 2 M sodium hydroxide (700 mL) was added. The ice bath was removed, and the reaction was stirred for 2 h to afford a clear brown 25 solution. The solution was acidified with 2 M sulfuric acid (400 mL) and stored at ca 5 0 C for 1 hour. A yellow solid formed and was collected by filtration. The solid was washed with water (3 x 100 mL) and recrystallized from 50% ethanol-water to afford compound 79 as tan needles (64.7 g, 82%).

57 Compounds 2-4, 6-17, 19-29, 34, 38-40, 42-48, 50-52, 67, 75-77, 89-94, 96, 100, 101, 107, 111, 116-118, 127-132, 134, 135, 193, 142, 143, 148, 149, 159, 162, 164, 169, 178-182, 187, and 190 were also prepared by this process.

Preparation of Compound 35. A solution of O-acetylsalicyloyl chloride (24.68 g, 124 mmol, 1 equiv) in tetrahydrofuran (300 mL) was cooled in an ice bath. Triethylamine (25 g, 249 mmol, 2 equiv) was added dropwise via an additional funnel. The methyl 9-aminononanoate hydrochloride was dissolved in DMF (190 mL, slightly warm to dissolve), charged to an addition funnel and added dropwise to the above mixture. The reaction was stirred in the ice-bath for 20 min and at room temperature for 2 h. Evaporation of the THF under reduced pressure gave a pink DMF solution. The pink solution was cooled in an ice-bath, and 2 M aqueous 15 sodium hydroxide (300 mL) was added. After being stirred at room temperature for 12 h, the mixture was acidified with 2 M hydrochloric acid (500 mL). The solution was cooled in an ice-bath, and a solid formed. The solid was collected by filtration and was recrystallized from ethanol/water to give compound 35 (32 g, 87%) as an off-white solid.

Preparation of Compound 49. 1-(2-hydroxyphenyl)-3-(4-methyl benzoate)-1,3-propane dione (3.00 g, 0.0101 mil.) is placed in a 100 ml round bottomed flask fitted with argon purge, magnetic stir bar and cold water condenser. Glacial acetic acid (20 mls) and concentrated sulfuric acid 25 (5 mis) were added, and heating of the reaction mixture was initiated. The reaction mixture was allowed to heat at reflux for 6 h before heating was discontinued. The reaction mixture was allowed to come to room temperature, and then was poured into 100 mis of ice/water. This was stirred for approximately 1/2 h before the mixture was filtered, and a brown -58solid was isolated. The brown solid was recrystallized twice from acetic acid, yielding compound 49 as a tan solid (1.44 g, 53.8%).

Preparation of Compound 167. 2-coumaranone (4.21 g, 0.0314 mol) was dissolved, with stirring, in acetonitrile (75 mis) in a 250 ml round bottomed flask fitted with a magnetic stir bar, argon purge and cold water condenser. Triethylamine (3.18 g, 0.0314 mol) and 8-aminocaprylic acid (5.00 g, 0.0314 mol) were added, and a tan slurry was formed. Heating was started, and the reaction mixture was allowed to reflux overnight. After heating overnight, thin layer chromatography of the reaction mixture ethyl acetate 50% hexane) indicated that the reaction had gone to completion. Heating was stopped, the reaction mixture was allowed to cool to room tempierature, and was concentrated in vacuo. The resulting residue was taken up in methylene chloride, and was washed with two, 100 ml 15 portions of 1N hydrochloric acid solution. The methylene chloride layer was dried with sodium sulfate and was concentrated in vacuo. The resulting tan *solid was allowed to dry in vacuo overnight, yielding compound 167 as a tan solid (8.35 g, 70.4%).

20 Preparation of Compound 171. 1,4-benzodioxan-2-one (3.93-g, 0.0262 mol) was dissolved, with stirring, in acetonitrile (70 mis) in a 250 ml round bottomed flask fitted with a magnetic stir bar, argon purge and cold o water condenser. Triethylamine (2.64 g, 0.0262 mol) and 8-aminocaprylic acid (500 g, 0.0262 mol) were added and a tan slurry was formed. Heating 25 was started, and the reaction mixture was allowed to reflux for approximately 3 hours. At this time, thin layer chromatography of the reaction mixture ethyl acetate /50% hexane) indicated that the reaction had gone to completion. Heating was discontinued, and the reaction mixture was allowed to cool to room temperature and was concentrated in vacuo. The resulting -59residue was taken up in methylene chloride and was washed with a 100 ml portion of 1N hydrochloric acid solution. At this time, a tan solid was noted to precipitate, and it was isolated by filtration. This tan solid was washed further with an additional 100 ml portion of 1 N hydrochloric acid solution, and then with 100 ml of water. The resulting tan solid was allowed to dry in vacuo overnight yielding Compound 171 as a tan solid (7.73 g, 95.6%).

Preparation of Compound 120. A solution of 3.00 g (18.3 mmol) of 2-nitrophenylisocyanate and 5 mL of tetrahydrofuran was dropwise over 10 min to an ice bath-cooled solution of 2.08 g (13.1 mmol) of 8aminocaprylic acid, 1.40 mL of 10 N NaOH and 40 mL of water. The reaction mixture was stirred an additional 30 min, warmed to 25 0 C and treated with 3% HCI solution until the pH was 5. The yellow precipitate was filtered off and rinsed with 100 ml of water. The yellow solid was 15 recrystallized in 2-propanol and water to give 3.7 g of compound 120 as pale yellow crystals.

Compounds 104-106 were also prepared by this procedure.

Preparation of Compound 133. A suspension of 2.40 g (16.3 20 mmol) and 2.80 g (15.6 mmol) of 4-(4aminophenyl)butyric acid in 20 mL-of propylene glycol, 2.40 mL (1.74 g, 17.3 mmol) of triethylamine and 10 mg (0.08 mmol) of dimethylaminopyridine was heated to 140°C. The mixture became a clear solution after 5 min at 1400C. After stirring for 330 min, the 0 reaction mixture was cooled to 25°C and diluted with 20 mL of water. The solid phthalimide which had formed was filtered off. The filtrate was acidified with 3% HCI solution. The resulting solid was filtered off and was recrystallized from 2-propanol and water to give 0.62 g of compound 133 as a tan solid.

Preparation of Compound 138. A solution of 1.73 g (12.9 mmol) of phthalic dialdehyde, 2.04 g 8-aminocaprylic acid and 20 mL of acetic acid was heated to reflux for 10 min. The reaction mixture was cooled to 40 0 C, diluted with water and extracted with CH 2

CI

2 (2 X 20 mL). The organic phase was washed with water and brine, dried over Na 2

SO

4 and evaporated. The residue was dissolved in ether and extracted with 2N NaOH.

The layers were separated. The aqueous layer was made acidic with 3% HCI and extracted with CH 2

CI

2 The organic phase was dried over Na 2

SO

4 and evaporated. The yellow residue was crystallized from acetonitrile and water to give 1.25 g of compound 138 as a yellow solid.

Preparation of Compound 140. A mixture of 1.40 g (9.48 mmol) 4 of phthalic anhydride and 1.51 g (9.48 mmol) of 8-aminocaprylic acid was heated to 150°C for 5 min. Upon cooling, 2.61 g of solid compound 140 S 15 was received.

Compound 150 was also prepared by this procedure.

Preparation of Compound 145. A suspension of 2.11 g (10.1 mmol) ethyl carbamoylanthranilic acid and 5 mL of CH 2 CIz was treated with 2.20 mL of oxalyl chloride. After stirring for 1 h the volatiles were stripped off. At that same time, a suspension of 1.60 g (10.1 mmol) of 8aminocaprylic acid and 15 mL of CH 2 CI, was treated with 2.60 mL (2.23 g, 20.5 mmol) of TMSCI. This mixture was heated to reflux for 90 min, cooled in an ice bath and treated with 4.30 mL (3.12 g, 30.9 mmol) of triethylarmine.

Five min later, a slurry of the residue from the oxalyl chloride reaction in mL of CH 2

CI

2 was added. The reaction mixture was warmed to 25 0 C and stirred overnight. Upon acidification of the mixture with 3% HCI, a white solid formed. The solid was filtered off and recrystallized from EtOH and water to give 1.88 g of compound 145.

-61 Compound 153 was also prepared by this procedure.

Preparation of Compound 154. A suspension of 4.02 g(25.6 mmol) of trans-4-aminomethylcyclohexane-carboxylic acid, 4.18 g (25.6 mmol) of isatoic anhydride, 20 mL of CH 2 CI,, 20 mL of dioxane, and 4 mL of water was heated to reflux for 12 h. The solution was cooled to 25 0 C and extracted with ether (4 x 20 mL). The organic layer was dried over NazSO, and concentrated. The resulting solid was recrystallized from EtOH and water to give 4.95 g of compound 154.

Compound 103 is available from Aldrich Chemical Company, Inc., Milwaukee, WI.

Example 2 Parathyroid Hormone Dosing Solutions Intracolonic dosing compositions containing 100 mg/kg of carrier and 25 pg/kg of parathyroid hormone in 25% aqueous propylene glycol or oral gavage dosing solution containing 400 mg/kg of carrier and 100 pg/kg of parathyroid hormone in water, were prepared with carriers 9, 33, 77, 79, 109, 110, 123, 136, 141, and 169. The dosing solutions are 00 designated P- carrier number

DS.

Comparative Example 2A Parathyroid Hormone Dosing Solutions An intracolonic dosing composition containing 100 mg/kg of a carrier having the formula 25

H

I

OH

-62and 25 ug/kg of parathyroid hormone in 25% aqueous propylene glycol was prepared. The dosing solution is identified as P-9A-DS.

Examples 3 In vivo Parathyroid Hormone Delivery Male Sprague-Dawley rats weighing between 200-250g were fasted for 24 hours and were administered ketamine (44 mg/kg) and chlorpromazine (1.5 mg/kg) 15 minutes prior to dosing. The rats were administered one of dosing solutions P-9-DS, P-33-DS, P-35-DS, P-77-DS, P- 79-DS, and P-141-DS by oral gavage or intra-colonic instillation Blood samples were collected serially from the tail artery for serum determination of parathyroid hormone concentration. Serum pardthyroid hormone concentrations were quantified by a parathyroid hormone immunoaccuracy test host.

Results are illustrated in Table 2, below.

Comparative Example 3A In vivo Parathyroid Hormone Delivery The procedure of Example 3 was followed substituting dosing solution P-9A-DS for dosing solution P-9-DS. Results are illustrated in Table 20 2, below.

Comparative Example 3B In vivo Parathyroid Hormone Delivery o 0 The procedure of Example 3 was followed with a dosing solution (at a dose of 25 pg/kg of parathyroid hormone (intra-colonic) or 100 pg/k-of 25 parathyroid hormone (oral)), P-0A-DS, that omitted the carrier.

•Results are illustrated in Table 2, below.

-63- TABLE 2 In vivo Parathyroid Hormone Delivery Mean Peak Serum (PTH] Dosing Solution Standard Deviation (pg/ml) P-9-DS 155 105 (IC) P-33-DS 58 18 (IC) 50 27 (IC) P-77-DS 358 274 (PO) P-79-DS 521 128 (PO) P-109-DS 128 25 (IC) P-110-DS 35 11 (IC) P-123-DS 49 22 (IC) P-136-DS 106 72 (IC) P-141-DS 120 120 (PO) P-169-DS 19 33 (IC) P-9A-DS 116 48 (IC) P-0A-DS 11 2 27 27 (IC) Examples 4 Recombinant Human Growth Hormone Dosing Solutions Intracolonic dosing compositions containing 25 mg/kg of carrier and 1 mg/kg of rHGH in phosphate buffer or oral gavage dosing solutions containing 600 5 mg/kg of carrier and 3 mg/kg of rHGH in phosphate buffer were prepared with carriers 9, 35, 36, 47, 62, 64, 67, 77, 79, 90, 94, 107, 109, 136, and J41.

The dosing solutions are designated R- carrier number DS.

Comoarative Example 4A Recombinant Human Growth Hormone Dosing Solutions -64- An intracolonic dosing solution was prepared according to the procedure of Example 4, substituting a carrier having the formula for the carrier. This dosing solution is designated as Comparative Example 4B Recombinant Human Growth Hormone Dosing Solutions An intracolonic dosing solution was prepared according to the procedure of Example 4, substituting a carrier having the formula

O

for the carrier. This dosing solution is designated as Comparative Example 4C Recombinant Human Growth Hormone Dosing Solutions An intracolonic dosing solution was prepared according to the procedure of Example 4, substituting a carrier having the formula 65

H

0 N

OH

for the carrier. This dosing solution is designated as R-9A-DS.

S 10 Example 5 In Vivo Recombinant Human Growth Hormone Delivery Male Sprague-Dawley rats weighing 200-250g were fasted for 24 hours and administered ketamine (44 mg/kg) and chlorpromazine mg/kg) 15 minutes prior to dosing. The rats were administered one of the dosing solutions of Example 3 by either oral gavage or intracolonic instillation.

Blood samples were collected serially from the tail artery for determination of serum rHGH concentrations. Serum rHGH concentrations were quantified by an rHGH immunoassay test kit.

Results are illustrated in Table 3, below.

Comparative Example 5A In Vivo Recombinant Human Growth Hormone Delivery The procedure of Example 5 was followed, substituting the dosing solutions of Comparative Examples 3A-3C for the dosing solutions.

Results are illustrated in Table 3, below.

Comparative Example 5B In Vivo Recombinant Human Growth Hormone Delivery The procedure of Example 5 was followed, with dosing solutions of active agent (at a dose of 1 mg of rHGH/kg (intracolonic) or 3 mg of -66rHGH/kg (oral) and no carrier. These dosing solutions are designated R-0D- DS and R-0E-DS, respectively. Results are illustrated in Table 3, below.

67 TABLE 3 In Vvo Recombinant Human Growth Hormone Delivery f Mean Peak Serum [rHGHJ ±t.

Dosing Solution j Standard Deviation (ng/ml) j R-9-DS 125 34 (IC) 41 46 (P0) 108 56 (IC R-36-DS 28 11 (IC) R-47-DS 0 (IC R-62-DS 11 12 (IC) R-64-DS 72 22 (P0) 19 22 (P0) -R-67-DS 88 24 (IC R-77-DS, 34 10 (P0) R-79-DS 62 51 (P0) 9 ±13 (P0) R-94-DS 39 35 (PO) R- 107-DS 0 0 (PO) R- 109-DS 128 ±25 (IC) R-1 36-DS 106 ±72 (IC) R-141-DS 95 14 (IC R-35A-DS 17 3 (IC) 42 28 (IC) R-9A-DS 55 17 (IC R-OD-DS 0 0 (IC) R-OE-DS 0 0 (IC -68- Example 6 In Vivo Interferon Delivery An intracolonic dosing composition containing 50 mg/kg of carrier 9 and 250 pg/kg of interferon in 50% propylene glycol was prepared.

Rats were administered the dosing composition by intracolonic instillation.

Delivery was evaluated by use of an ELISA assay for human interferon a from Biosource, Inc. Mean peak serum interferon concentration was 2611 695.

Comparative Example 6A In Vivo Interferon Delivery Rats were administered, orally and by intracolonic instillation, dosing solutions of 1 mg/kg of interferon and no carrier. Delivery was evaluated according to the procedure of Example 6. Mean peak serum interferon concentration was 1951 1857 (PO) and 79 100 (IC).

Example 7 Heparin Dosing Solutions Intracolonic dosing compositions containing 50 mg/kg of carrier and 25 mg/kg of heparin in 25% aqueous propylene glycol or oral gavage dosing solutions containing 300 mg/kg of carrier and 100 mg/kg of heparin in 25% aqueous propylene glycol were prepared with carriers 9, 35, 47, 58, 62, 64, 67, 76, 96, 102, 109, 110, 111, 117, 122, 123, 139, 141, 144, 20 and 169. The dosing solutions are designated H-carrier number-DS.

Comparative Example 7A Heoarin Dosing Solutions Comparative intracolonic dosing compositions were prepared according to the procedure of Example 7, substituting the following carriers 25 for the carrier.

0 0 nN ~0 0 A -69-

OH

0 o These dosing solutions are designated H-35A-DS, H-35B-DS, and H-109A-DS, respectively.

Examples 8 In Vivo Evaluation of Heparin in Rats The dosing solutions of Example 7 were administered to fasted rats either by oral gavage or intracolonic instillation.

Blood samples were collected by cardiac puncture following the 20 administration of ketamine (44 mg/kg). Heparin activity was determined-by utilizing the activated partial thromboplastin time (APTT) according to the method of Henry, Clinical Diagnosis and Management by Laboratory Methods; Philadelphia, PA; W.B. Saunders (1979).

Results are in illustrated in Table 4, below.

•Comparative Examples 8A In Vivo Evaluation of Heparin in Rats The dosing solutions of Comparative Example 7A were administered to fasted rats by intracolonic instillation. Blood samples were collected and heparin activity was determined by the method of Example 8.

collected and heparin activity was determined by the method of Example 8.

1 Results are illustrated in Table 4, below.

Comparative Example 8B In Vivo Evaluation of Heparin in Rats An intracolonic dosing solution of 25 mg/kg of heparin and an oral gavage dosing solution of 100 mg/kg of heparin were administered to fasted rats. These dosage solutions were designated H-0A-DS and H-0B-DS, respectively.

Blood samples were collected, and heparin activity was determined by the methods of Example 8.

Results are illustrated in Table 4, below.

*I

71 I TABLE 4 In Viva Evaluation of Heparin in Rats Dosing Solution JHeparin APTT (sec) H-9-DS 48 18 (IC) 54 27 177 85 (I) H-47-DS 30 14 (IC) 40 22 (IC) H-58-DS24 4 (IC) H-62-DS37 13 (IC) H-6-DS59 28 168 ±75 IC) H-67-DS76 36 (IC) H-76-DS 63 27 (PO) H-96-DS36±8(C 102-DS il±18c H-1 09-OS 5 8O H-i 10 -OS37±9O H-ill -OS 7 9P H-1i17-DS 140 128 (IC H-122-DS 49 21 207 ±7 (PO) H- 123-DS 42 14 (PO) H-139-DS 31 I I C) H-141-DS 59 26 (IC) H- I44-DS 26 3 (IC) H-35A-DS 61 29 (IC) H-358-DS 51 30 (IC) H-169-DS 23 2 PIC H-OA-DS 23 2 (P0) H-08-DS 33 6 (IC) 9 e 0 0a 0 0 0 .5ee P:\WPDOCSCABhSPEC\76290.d-O2/V02 72 The above mentioned patents, applications, test methods, and publications are hereby incorporated by reference in their entirety.

Many variations of the present invention will suggest themselves to those skilled in the art in light of the above detailed description. All such obvious variations are within the full intended scope of the appended claims.

The reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that that prior art forms part of the common general knowledge in Australia.

Amendments have been made to the claims which follow from the claim set as originally filed in parent application No. 62756/98 to more closely define the invention.

The claim amendments are based on various preferred embodiments as described and exemplified in the description. However, for the purpose of maintaining integrity of the text of the disclosure, substantial amendments have not been made to the description in light of the claim amendments.

oooeo o*

Claims (11)

1. 2. The composition of claim 1, wherein the active agent is selected from the group consisting of biologically active agents, chemically active agents, and any combination thereof.
2. 3. The composition of claim 2, wherein the biologically active agent comprises at least one peptide, mucopolysaccharide, carbohydrate, or lipid.
3. 4. The composition of claim 2, wherein the biologically active agent is selected from the group consisting of human growth hormones, bovine growth hormones, growth hormone-releasing hormones, interferons, interleukin-1, interleukin-II, insulin, heparin, low molecular weight heparin, calcitonin, erythropoietin, atrial naturetic factor, antigens, monoclonal antibodies, somatostatin, adrenocorticotropin, gonadrotropin releasing hormone, oxytocin, vasopressin, cromolyn sodium, vancomycin, desferrioxamine, •parathyroid hormone, and any combination thereof. The composition of claim 4, wherein the biologically active agent comprises interferon, interleukin-1, interleukin-II, insulin, heparin, low molecular weight heparin, calcitonin, oxytocin, vasopressin, vancomycin, desferrioxamine, pgrathyroid hormone, or CA\TEMME.WN, I# 109A=- IV) 2AMn 0 1 -74- any combination thereof.
4. 6. The composition of claim 5, wherein said biologically active agent comprises calcitonin.
5. 7. A dosage unit form comprising the composition of any of claims 1 to 6; and an excipient, a diluent, a disintegrant, a lubricant, a plasticizer, a colorant, a dose vehicle, or any combination thereof.
6. 8. The dosage unit form of claim 7, comprising a tablet, a capsule, or a liquid.
7. 9. The dosage unit form of any of claims 7 and 8, wherein said dosing vehicle is selected from the group consisting of water, 1,2-propane diol, ethanol, and any combination thereof.
8. 10. A method for administering a biologically-active agent to an animal in need of said agent, said method comprising administering orally, intranasally, sublingually, intraduodenally, subcutaneously, rectally, vaginally, bucally or ophthalmically to said animal the composition of any of claims 1 to 6. Fq)TEMMEmi-DivINIlo ,.doc-1112AX) I
9. 11. A compound having the formula OH O N SH 0 CI or a salt thereof.
10. 12. A method for preparing a composition, said method comprising mixing: at least one active agent; the compound of claim 11; and optionally, a dosing vehicle.
11. 13. A method for passing a biologically active agent across the blood/brain barrier of an animal in need of said agent, said method comprising administering to said animal the composition of any of claims 1 to 6. S14. A composition comprising calcitonin; and a compound having the formula OH O OHOH ^^Nt or a salt thereof. :pP:kWPDOCSH jwXSpcsAEnis-Div 11 AW- I /12l( -76- Method for the manufacture of a compound of claim 11, compositions containing said compound and/or uses of said compound substantially as hereinbefore described with reference to the Examples. DATED this 14th day of December 2000 Emisphere Technologies, Inc. By its Patent Attorneys DAVIES COLLISON CAVE 000e *9a