BRPI0719319A2 - COLONIC RELEASE USING ZN / PECTIN ACCOUNTS WITH AN EUDRAGIT COATING - Google Patents

Description

Patent Descriptive Report for "COLONIC RELEASE USING Zn / PECTIN ACCOUNTS WITH A EUDRAGIT COATING".

Field of the Invention

The present invention is in the field of oral drug delivery systems for delivering active agents, such as metallospecific enzymes, to the colon.

Background of the Invention

Drug delivery systems that specifically release active agents into the colon have been recognized as having important therapeutic advantages. A large number of colonic conditions could be effectively treated more effectively if the active ingredient were locally released. Examples of such colonic disorders include Crohn's disease, ulcerative colitis, colorectal cancer and constipation.

Colonic release may also benefit patients when, from a therapeutic point of view, a delay in absorption is required. Examples include treatment of disorders such as nocturnal asthma or angina (Kinget R. et al. (1998), Colonic Drug Targeting, Journal of Drug Targeting, 6, 129).

Colonic release may also be used to administer therapeutically active polypeptides. Polypeptides are typically administered by injection because they are degraded in the stomach. Because injection is painful, investigative efforts have focused on using the colon as an absorption site for active polypeptides, including painkillers, preservatives, vaccines, insulin, and the like. Absorption of polypeptides in the colon appears to be more effective than elsewhere in the digestive tract. This is particularly due to the relatively weak proteolytic activity in the small intestine and the absence of membrane-associated peptidase activity of colonic epithelial cells.

Following their oral administration, antibiotics cross the stomach and are then absorbed into the small intestine to spread throughout the body and treat the infectious eruption site (s) to which they were administered. Also, an ingested fraction of antibiotics (the importance of this fraction varies with the characteristics of each antibiotic) is not absorbed and continues its progress to the colon before it is eliminated in the stool. These residual antibiotics are combined in the large intestine with a fraction of the absorbed antibiotics but which are excreted in the digestive tract by biliary elimination. This fraction is of varying importance as a function of metabolism and elimination pathways for each antibiotic. Finally, for some antibiotics, a fraction of the absorbed dose is excreted directly from the blood through the intestinal mucosa back into the digestive tract lumen, a good example is known as cyprofoxacin. Thus, if administered orally or parenterally, a residual fraction of the active antibiotics is usually found in the colon. This is the case, to varying degrees, for the vast majority of antibiotics of the various families used in therapy, with the notable exception only of the aminoglycoside antibiotics for which intestinal excretion is insignificant. For other antibiotics, intestinal excretion of residual antibiotic activity will have a variety of consequences, all detrimental. In fact, the colon is home to a complex and very dense bacterial ecosystem (several hundred different bacterial species; more than 1011 bacteria 20 per gram of colonic content) that will be affected by the arrival of active antibiotic residues. The following can be observed:

1. Flora imbalance that is the leading cause of banal diarrhea occurring following antibiotic treatments (Bartlett J. G. (2002) Clinical practice. Antibiotic associated diarrhea, / Vew England Journal of Medicine,

346, 334). Although this diarrhea is usually non-serious and ceases rapidly, either spontaneously or upon completion of antibiotic treatment, it is adversely perceived by patients and adds to the discomfort of the original disease for which the antibiotic was prescribed;

2. Interference with resistance to colonization through exogenous bacteria (or "barrier effect") with possible risk of infection, such as

as salmonella food poisoning (Holmberg S. D. et al. (1984) Drug resistant Salmonella from fed antimicrobial animals, New England Journal of Medicine, 311,617);

3. Selection of antibiotic resistant microorganisms. These microorganisms can be of several types:

a) first they may be pathogenic bacteria such as for example Clostridium difficile, a species capable of secreting toxins that

cause a form of colitis known as pseudomembranous colitis (Bartlett J. G. (1997) Clostridium difficile infection: pathophysiology and diagnoses, Seminar in Gastrointestinal Disease, 8, 12);

b) they may also be microorganisms that are relatively weak pathogens but whose multiplication may lead to infection

(Vaginal Candidosis or Escherichia coli-resistant cystitis).

c) they may ultimately be bacteria resistant to non-pathogenic co-monthly drug whose multiplication and fecal elimination will increase the spread of antibiotic resistance in the environment. It's well documented

Since antibiotic resistance genes are carried by mobile or transposable genetic elements that can contain up to 5 or 6 antibiotic resistance genes, and are easily transmitted to other bacteria, even across species. Therefore, these resistant commensal bacteria may be an important source of drug resistance for pathogenic species. This risk is currently considered seminal in terms of the restlessness of the evolution to drug resistance by numerous pathogenic species for humans.

It would therefore be desirable to have drugs and drug delivery systems that would act to reduce the amount of residual antibiotics reaching the colon following oral or parenteral antibiotic therapy.

Numerous strategies that explore the various physiological parameters of the digestive tract have been invented with the goal of releasing active ingredients in the colon. These strategies focused on drug release systems based on (1) using polymers that are sensitive to pH variations, (2) time-dependent drug release forms, (3) prodrugs or bacterial degrading polymers. intestinal flora.

It would be advantageous to have additional drug delivery systems that allow the administration of active agents to the colon, including, but not limited to, agents that reduce the amount of residual antibiotics in the colon. The present invention provides such drug delivery systems. Summary of the Invention

Drug delivery systems that can release prophylactic, therapeutic and / or diagnostic agents to the colon are disclosed. Systems include cross-linked pectin beads, for example with a metal zinc cation or any divalent cation of interest whose beads are then coated with Eudragit® type polymers.

Other embodiments are set forth in the claims which are incorporated herein in their entirety by reference.

Drug delivery systems are orally administrable, but may release active agents to the colon. In some embodiments, they may administer the agents at various positions in the gastrointestinal tract, including the colon.

In one embodiment, the therapeutic agent is an agent capable of reducing the residual amount of residual antibiotics that reach the

120 colon following oral or parenteral antibiotic therapy, such as a metallo-dependent enzyme. Application is illustrated for Stenotropophomonas maltophilia β-lactamase L1. However, non-metalloenzyme β-lactamases (classes A, C or D) may also be used. In addition, enzymes, metallo-dependent or not, may be used to inactivate other classes of antibiotics such as macrolides, quinolones and fluoriquinolones, glycopeptides, lipeptides, cyclins, oxazolidinones, and other classes of antibiotics. Enzymes may have the full sequence of the native enzyme, or may be truncated or otherwise modified as long as they maintain acceptable activity. Release agents capable of reducing the amount of residual antibiotics that reach the colon following oral or parenteral antibiotic therapy may limit the development of bacterial resistance. In other embodiments, therapeutic agents include, but are not limited to: • Natural peptides and proteins (including, but not limited to, enzymes, hormones, cytokines, lymphokines, growth factors, antibodies, and the like) synthetic or recombinant;

• Nucleic acids and compounds including 5 nucleic acid elements (including, but not limited to, plasmids, oligonucleotides (o-

ligoribonucleotides, deoxyribonucleotides, SiRNAs or ShRNAs of various lengths, and mixed molecules, including natural and / or modified bases, and optionally containing substitutions and modifications), as well as peptide nucleic acids;

· Complex structures of natural, recombinant or synthetic origin

including but not limited to viruses (including DNA and RNA viruses, animal cell-marking viruses, plant cell-marking viruses, or viruses that mark bacteria better known as bacteriophages), bacteria (in any including spores), mycoplasmas, yeast and other

other unicellular eukaryotes (in any form including spores)

• Natural, synthetic or mixed chemical molecules or mixtures thereof of any size, class or structure;

• Compounds for use in the diagnosis, treatment or investigation of humans and animals for any reason or condition, including diseases.

infectious diseases (including but not limited to those of bacterial and viral origin), inflammatory diseases, cancers;

• Compounds to aid, supplement or modify treatment with anti-infectious, anti-inflammatory, anticancer, immunomodifying agents, and others, particularly where such aid,

Implementation, or modification, refers to the ability to block or modulate receptor activity in the colon, or inactivate other therapeutic agents that could modulate receptor activity in the colon.

Colon-specific release is obtained by formulating a prophylactic, therapeutic, and / or diagnostic agent, such as a meta-enzyme.

dependent drug or other agent capable of reducing the amount of residual antibiotics that reach the colon following oral or parenteral antibiotic therapy with specific polymers that degrade in the colon, such as pectin. Pectin is gelled / crosslinked with a cation such as a zinc cation. The formulation, typically in the form of ionically crosslinked pectin beads, is subsequently coated with a specific polymer such as an Eudragit® polymer.

Release may be modulated to occur at various sites pre-

selected from release into the intestinal tract by gelling / cross-linking a mixture of the prophylactic, therapeutic, and / or diagnostic agent and pectin with divalent metal cations such as Ca2 + or Zn2 +.

10 Past Efforts Focused on Coating Pectin Beads

with cationic polymers such as polyethylene imine (PEI), chitosan or other cationic polymers to prevent pectin beads from degrading in the upper gastrointestinal tract. Such efforts are described, for example, in U.S. Patent Application 10 / 524,318, and U.S. Patent Application.

No. 60 / 651,352, the contents thereof are hereby incorporated by reference.

The present invention relates to the coating of pectin beads with Eudragit® polymers such as FS30D, L30D (also known as L30D-55), NE30D, mixtures thereof or other types thereof.

* 20 targetable Eudragit® polymers to achieve desired release of prophylactic, therapeutic and / or diagnostic agent at predefined levels of the gastrointestinal tract (GIT).

When the Eudragit® coating is dissolved according to certain parameters such as pH or time, the beads are preferably degraded by pectinolytic enzymes found in the lower intestinal tract. Pectin degradation then releases the prophylactic, therapeutic and / or diagnostic agent encapsulated within the bead.

One aspect of the invention is to provide a stable metalloenzyme formulation for the lower intestinal or colonic release of such an enzyme. The use of zinc cations to crosslink pectin is particularly preferred when specific metallo-dependent enzymes, which are Zn2 + -dependent, can interact with other cationic species if they are used to gel pectin beads. Such interactions could drastically affect the activity of such metallo-dependent enzymes. Accordingly, one embodiment of the drug delivery system involves using Zn2 + ions as a cross-linking agent for 5 pectin beads and in association with Zn + -dependent enzymes which are very sensitive to the presence of other competitive cations. Of course, if the enzymes are dependent on other metal cations, such other metal cations (if they have a valence exceeding +1) can be used to crosslink the pectin.

The processes for obtaining such accounts may involve conditions

specific procedural conditions, such as time for gelation, washing, and drying, which can be optimized to provide higher quality beads with optimized in vitro and in vivo efficacy. Therefore, another embodiment of the invention relates to processes for preparing zinc crosslinked and Eudragit® coated pectin beads.

Brief Description of the Figures

Figure 1 is a graph showing the efficiency of water rinsing to remove excess metal cations from a β-Iacatamase L1 formulation on zinc acetate crosslinked pectin beads measured in conductivity (mS / cm) per sample following several washings.

Figure 2 is a graph showing the effect of gelling time, rinsing process, and drying time upon restoration of β-lactamase L1 activity.

Figure 3 is a graph showing the enzymatic activity of β-

L1 lactamase using CENTA as a substrate, measured in response (OD / min) versus L1 concentration (pg / ml).

Figure 4 is a series of scanning electron micrographs showing Eudragit-coated beads prepared using the methods described herein, and a cross-section of the beads showing the approximate thickness of the Eudragit layer.

Figure 5 is a table showing the release kinetics of β- * Lactamase L1 from uncoated beads, and Eudragit coated beads with or without hydroxypropyl methyl cellulose (HPMC) precoated, measured in activity (pg / mg of bills) versus time (minutes). Blue triangles represent uncoated beads; red circles represent beads coated with 40% Eudragit L30D-55 without pre-coating; Green squares represent 5% HPMC pre-coated beads and 40% Eudragit L30D-55 coated beads.

Figure 6 is a table showing amoxicillin hydrolysis by uncoated, Eudragit coated beads with or without a pre-coated hydroxypropyl methyl cellulose (HPMC), measured in terms of residual amoxicillin (%) versus time (minutes) . Blue triangles represent uncoated beads; red circles represent beads coated with 40% Eudragit L30D-55 without pre-coating; green squares represent HPMC pre-coated and Eudra-15 git L30D-55 coated beads.

Figure 7 is a table showing the effect of L1 β-lactamase-containing Eudragit coated pectin beads on emergence of amoxicillin-treated antibiotic resistant bacteria, measured in terms of amoxicillin-resistant bacteria (%) versus trajectory duration -> 20 treatment (days). Blue triangles represent untreated animals (n = 12); red lozenges represent amoxicillin-treated animals and placebo pectin beads (n = 12); green squares represent amoxicillin-treated animals along with Eudragit-coated pectin beads containing β-lactamase L1 (n = 4).

Detailed Description of the Invention

The drug delivery systems described herein will be better understood with reference to the following detailed description.

I. PECTIN ACCOUNTS

Pectin beads are formed of pectin, zinc ions, and other coating with Eudragit® polymers and encapsulated with one or more active agents.

Stability and protection of pectin beads in gastric and intestinal media are ensured by the Eudragit ® polymer coating. In contrast, non-pectin-coated beads tend not to be stable in such an environment and may not adequately protect their contents against degradation and / or inactivation. The Eudragit ® 5 coating ensures that they are durable enough that their contents are able to reach the colon intact.

Pectin

Pectin is a polysaccharide isolated from the upper plant cell walls, widely used in the agricultural food industry (such as a clot or thickener for jellies, ice cream and others) and pharmaceutical science. It is polymolecular and polydispersed. Its composition varies depending on the source, extraction conditions and environmental factors. Pectins are mainly composed of linear chains of beta-1,4- (D) -galacturonic acid, sometimes interspersed with rhamnose units. The carboxylic groups of galacturonic acid may be partially esterified to yield methylated pectins. Two types of pectins are distinguished according to their degree of methylation (DM: number of methoxy groups per 100 units of galacturonic acid):

- highly methylated pectin (HM: high methoxy) where the degree of methylation ranges from 50 to 80%. It is slightly soluble in water and forms

gels in acidic medium (pH <3,6) or in the presence of sugars;

- weak methylated pectin (LM: low methoxy), with a degree of methylation ranging from 25 to 50%. More soluble in water than pectin HM, it gives gels in the presence of divalent cations such as Ca2 + ions. In fact, the

Ca2 + ions form "bridges" between the free carboxylated groups of the galacturonic acid halves. The network that is formed has been described by Grant et al. under the name 'egg box model' (Grant G. T. et al. (1973) Biological interactions between polysaccharides and divalent cations: the egg-box model, FEBS Letters, 32, 195).

There are also amidated pectins. Treatment of pectin by ammonia

It does not transform some methyl carboxylate groups (-COOCH 3) into carboxamide groups (-CONH 2). This amidation imparts new properties to the tetras, in particular better resistance to variations in pH. Amide pectins tend to be more tolerant to pH variations and have been studied for the manufacture of colonic release matrix tablets (Waker-Iy Z. et al. (1997). Studies on amidated pectins as potential carriers in colonic drug delivery , Journal of Pharmacy and Pharmacology (49, 622).

Pectin is degraded by enzymes originating from higher plants and various microorganisms (fungi, bacteria, and the like) including bacteria from the human colonic flora. The enzymes produced by microflora comprise a mixture of polysaccharidases, glycosidases and sterols.

Zinc Cations

Divalent zinc cations can be used from various zinc salts to crosslink pectin. Examples include zinc sulfate, zinc chloride, and zinc acetate.

In the present invention, enteric coating agents

are preferably alkyl methacrylic acid-acrylate copolymers such as Eudragit® polymers.

Eudragit® Polymers

Coating of drug-loaded cores, such as 20 tablets, capsules, granules, pellets or crystals, offers many advantages over uncoated counterparts, such as higher physicochemical stability, improved compliance and increased therapeutic efficiency of ingredients. active. In fact, the effectiveness of a medication depends not only on the assets it contains, but also on the formulation and processing.

Poly (meth) acrylates have proven particularly suitable as

coating materials. These polymers, typically used in amounts of only a few milligrams, are pharmacologically inactive, ie excreted unchanged.

EUDRAGIT® is the trade name for copolymers derived from acrylic and methacrylic acid esters whose properties are determined by functional groups. Individual EUDRAGIT® types differ in their proportion of neutral, alkaline or acidic groups and thus in terms of physicochemical properties. The skillful use and combination of different EUDRAGIT® polymers offer ideal solutions for controlled drug release in various pharmaceutical and technical applications. EU-DRAGIT® provides functional continuous release tablet films and 5 pellet coatings. Polymers are described in international pharmacopoeias such as pH. Eur., USP / NF, DMF and JPE.

EUDRAGIT® polymers may provide the following possibilities for controlled drug release:

• Targeting of the gastrointestinal tract (gastroresistance, Colon clearance)

• Protective coatings (taste and odor masking, moisture protection)

• Drug release (continuous release formulations).

EUDRAGIT® polymers are available in a wide range of

of different concentrations and physical forms, including aqueous solutions, aqueous dispersion, organic solutions, and solids.

The pharmaceutical properties of EUDRAGIT® polymers are determined by the chemical properties of their functional groups. A distinction is made between:

· Poly (meth) acrylates, soluble in digestive fluids (through

salt formation)

EUDRAGIT® L (Methacrylic Acid Copolymer), S (Methacrylic Acid Copolymer), FS and E (Basic Butyl Methacrylate Copolymer) polymers with acidic or alkaline groups allow pH-dependent release of the active ingredient.

Applications: Simple taste masking only through gastric fluid resistance, for controlled drug release in all sections of the intestine.

• Poly (meth) acrylates, insoluble in digestive fluids EUDRAGIT® RL and RS polymers (methacrylate copolymers)

of ammonia) with alkaline polymers and EUDRAGIT® NE with neutral groups allow controlled time release of the active through pH-independent swelling.

Information about Eudragit polymers is found at: http: //www.pharma-

polymers.com/pharmapolymers/en/eudragit/entericcoatings/ and at 5 http: //www.pharma-

polimers.com/pharmapolymers/en/eudragit/regulatorytoxicology/ Enteric Coatings: Gastroresistance and Colon Release

EUDRAGIT® enteric coatings provide protection against drug release in the stomach and allow controlled release in the intestine. Marked drug release in the gastrointestinal tract is recommended for particular applications or therapeutic strategies, for example when the drug is poorly soluble in the upper digestive tract, or when the drug may be degraded by the gastric fluid. Secondly, this dosage form is very patient friendly as it does not strain the stomach and the number of doses of the therapeutic drug can be considerably reduced thanks to the prolonged release. The dominant criterion for release is the pH-dependent dissolution of the lining that occurs in a certain section of the intestine (pH 5 to more than 7) rather than the stomach (pH 1-5). For these applications, EUDRAGIT® anionic type containing> ■ 20 carboxyl groups can be mixed together. This makes it possible to finely adjust the dissolution pH, and thereby define the drug release site in the gut. EUDRAGIT® types LeS are suitable for enteric coatings. EUDRAGIT® FS 30 D (aqueous dispersion of an anionic copolymer based on methyl acrylate, methyl methacrylate and methacrylic acid) is specifically used for controlled release in the colon.

Benefits of applying endemic EUDRAGIT® coatings include:

• pH-dependent drug release

• protection of gastric fluid sensitive assets

30 · gastric mucosa protection from aggressive assets

• increased drug effectiveness

• good storage stability • controlled release in colon bleaching / GI Active Agents

The active agent may be an anti-infectious, for example antibiotics, anti-inflammatory compounds, antihistamines, anticholinergics, antivirals, antimitotics, peptides, proteins, enzymes, nucleic acids (RNA or DNA), nucleic acids. peptides, plasmids, genes, antisense oligonucleotides, interfering RNA, ribozymes, small molecules with specific binding capacities or activities (such as labeled chemotherapeutic), diagnostic agents, immunosuppressive agents, viruses, bacteria, other microorganisms eukaryotic organisms or cells.

The active agent may be introduced into the drug delivery system as a powder, solution, suspension, or complexed with a solubilizing agent such as a cyclodextrin or any other suitable compound.

Some of the active agents described herein may be administered by

in the form of prodrugs. Prodrugs have been widely studied for the colonic bleaching of various active ingredients (such as steroidal and non-steroidal anti-inflammatory drugs, and spasmolytic drugs).

These systems are based on the ability of the enzymes pro-

induced by the colonic flora to act on prodrugs to release the active form of the active ingredient.

Prodrugs can be based on the action of bacterial azoductases, so that active agents are labeled at the 25 Ion with the drug release systems described here, and active agents are formed by reaction of the prodrug. A drug with a bacterial azoreduct that provides a dual mechanism to ensure that drugs are administered to the colon. Representative chemistry for forming such prodrugs is described, for example, in Peppercorn Μ. A. et al. (1972) 30 The role of intestinal bacteria in the metabolism of salicylazosulfapyridin, The Journal of Pharmacology and Experimental Therapeutics, 181, 555 and 64, 240. * · Another method is to use bacterial hydrolases such as

glycosidases and polysaccharidases (Friend DR (1995) Glycoside prodrugs: novel pharmacotherapy for colonic diseases, STP Pharma Sciences, 5, 70; Friend DR et al. (1984) A colon-specific drug delivery system based on 5 drug glycosides and the glycosidases of colonic bacteria, Journal of Medicinal Chemistry, 27, 261; Friend DR et al. (1985) Drug glycosides: potential prodrugs for colon-specific drug delivery, Journal of Medicinal Chemistry, 28, 51; and Friend DR et al. (1992) Drug glycosides in oral colon-specific drug delivery, Journal of Controlled Release, 19, 109). Prodrugs have thus been developed by coupling, for example, sugar with steroids (glucose, galactose, cellobiose, dextran (international patent application WO 90/09168)), cyclodextrins Hirayama F. et al. (1996) In vitro evaluation of Biphenyly Acetic Acid-beta-Cyclodextrin conjugates to colon-targeting drugs: drug release behavior in rat biological media, Journal of Pharmacy 15 and Pharmacology, 48,27).

a) Antibiotic Inactivating Agents

In one embodiment, the active agent is an enzyme capable of inactivating antibiotics in the colon. Any agent that inactivates an antibiotic can be administered.

When the antibiotic is a beta-lactam antibiotic, β-

Iactamases may be used. The selected enzyme, ie β-lactamase L1, a Stenotrophomonas maltophilia β-lactamase Zn2 + -dependent, was chosen from a series of β-lactamases because its characteristics showed the best profile for the marked application. Also, it has been shown to have an excellent stability profile. The characteristics of the various β-Iaetamases evaluated are described hereafter.

There are a variety of enzymes that are known to be metallo-dependent, in addition to the β-lactamase L1 enzyme discussed above. When it is desired to administer such enzymes to a patient by oral administration, care should be taken to avoid having the enzyme digested in the stomach or upper intestine. Accordingly, the drug delivery system described herein can be advantageously used to release such metallo-dependent enzymes. The cation used to crosslink pectin comprises the cation on which the enzyme depends.

A representative enzyme is β-lactamase L1, a Zn2 + -dependent Stenotrophomonas maltophilia β-lactamase, which was chosen from a series of β-lactamases because its characteristics showed the best profile for the marked application. Also, it has been shown to have an excellent stability profile. The characteristics of the various β-lactamases evaluated are described hereinafter.

When the antibiotic is from another class of antibiotics, enzymes or other molecules that inactivate such antibiotics may be used. One such example would be to use an erythromycin esterase to inactivate macrolide antibiotics.

A representative erythromycin esterase is that disclosed by Andremont A. et al. ((1985) "Plasmid mediated susceptibility to intestinal 15 microbial antagonisms in Escherichia coli", Infect. Immun. 49 (3): 751), the contents thereof are hereby incorporated by reference. When the antibiotic is a quinolone, the active agent may be one capable of inactivating the quinolones. Representative agents include those disclosed by Chen, Y et al. ((1997) "Microbicidal models of soil metabolism biotransformations of 20 danofioxacin", Journal of Industrial Microbiology and Biotechnology 19: 378).

Patients may be treated with combinations of these

people.

Representative β-lactamase enzymes, and their efficacy in various antibiotics, are shown in Table 1.

Table 1

Antibiotics FEZ-1 * Km (MM) Kcat / K ™ L-1 (p) ** Km KcatZKm Kcat (S_1) (μΜ / S'1) Kcat (S-1) (μΜ) (μΜ / S'1) Penicillin Benzylpenicillin 70 590 0.11 600 38 16 Ampicillin> 5.5> 5000 0.011 520 55 9.5 Cabenicillin 35 1600 0.023 Continued ...

Pipericillin 50 4200 0.012 Azloeiline Mezloeiline Thiearciline> 65> 5000 0.013 Thymoiline Cephalosporin Cephaloridine 16 1000 0.016 Cephalothin 300 120 2.5 82 8.9 9.2 Cefoxoxin 1 3 0.33 Cefuroxin Cefotaxim 165 70 2.4 270 10 27 Ceftazidine Cefazidine 6> 1000 0.006 Cefpirom Nitroeeffin 90 100 0.9 41 4 10 Moxalaetama 3 18 0.17 Carbapenem Imipenem> 200> 1000 0.2 370 57 6.5 Meropenem 45 85 0.5 157 15 10 Biapenem 70> 1000 0.07 134 32 4.2 Aztreonam Carumonam Monobactams Mechanically based inactivators Sulbaetama Tazobaetama 40 700 1.06 Clavulanic acid <0.01> 1000 <0.000 11 22 0.5 01 Penicillin Benzylpenieiline 320 520 0, 62 56 49 1,114 Ampieillin 950 200 4,8 125 90 1,4 I Cabenicillin 0,02 185 205 0,9 Pipericillin 0,72 300 125 2,4 Azlocillin 200 200 1 Mezloeiline 200 125 1,4 Thioureylin 1,1 740 0 , 0015 180 125 1.6 Thymoeiline> 2000 <0.0001 7.7 390 0.002 Cephalosporin Cephaloridine 53 22 2.4 140 50 2.8 Cephalothin 48 21 2.4 130 11 12 Cefoxitin 16 8 2 15 13 1.2 Cefuroxin 8 37 0, 22 8 20 0.4 Cefotaxim 1.3 4 0.45 70 12 5.8 Ceftazidine 8 44 0.18 3.6 72 0.05 Cefepim 7 11 0.66> 40> 400 0.1 Cefpirom 9 14 0, 64 180 180 1 Nitroeeffin 63 27 2.3 770 18 43 Moxalaetam 90 55 1.6 Carbapenem Imipenem 46 39 1.2 34 9 3.8 Meropenem 50 10 5 2 2.5 Biapenem 160 28 6 8.5 15 0.55 Aztreonam Monobactams> 0.01 > 1000 <0.0001 <0.01> 1000 <0.000 1 Carumonam> 0.01> 1000 <0.0001 Mechanism-Based Inactivators Sulbaetam 23 320 0.072 Tazobaetam 28 875 0.032 Lavulanic Acid * (Mercuri et al, Antimicrob . Agents. Chemother. April 2001; 45 (4): 1254-1262)

** (Carenbauer et al., BMC Biochem. 2002; 3: 4. Epub February 2002. 13; Frere1 2005, unpublished data)

*** (Murphy et al, 2003, Antimicrob. Ag. Chemother. February 2003, 47 (2): 582-7; Laraki et al., Antimicrob. Ag. Chemother. April 1999, 43 (4): 902 -6)

5 **** (Docquier et al., J Antimicrob. Chemother. February 2003, 51 (2): 257-266)

b) Agents Treating Colon Cancer

When drug delivery systems are used to treat colon cancer, any type of antitumor agent may be used. The anti-tumor agents may be, for example, antiproliferative agents, DNA modification or repair agents, DNA synthesis inhibitors, DNA / RNA transcription regulators, RNA processing inhibitors, agents that affect the protein expression, synthesis and stability, agents that affect protein localization or its ability to exert its physiological action, agents that interfere with protein-protein or protein-nucleic acid interactions, agents that act through RNA interference, receptor binding molecules of any chemical nature (including small molecules and antibodies), labeled toxins, enzyme activators, enzyme inhibitors, gene regulators, HSP-90 inhibitors,

1 20 molecules that interfere with microtubules or other cytoskeletal components or cell adhesion and motility, phototherapy agents, and therapy aids.

Representative antiproliferative agents include N-acetyl-D-sphingosine (C2 ceramide), apigenin, berberine chloride, dichloromethylenodiphosphonic acid disodium salt, loe-emodine, emodin, HA 14-1, N-hexanoyl-D-sphingosine (ceramide C6), 7b-hydroxycholesterol, 25-hydroxycholesterol, hyperforin, parthenolide, and rapamycin.

Representative DNA modification and repair agents include aphidicoline, bleomycin sulfate, carboplatin, carmustine, chlorambutyl, cyclophosphamide monohydrate, ISOP AC® cyclophosphamide monohydrate, cis-diaminoplatin dichloride (ll) (cystplatin), esculetin , melfalan, methoxyamine hydrochloride, mitomycin C, mitoxantrone dihydrochloride, oxaliplatine, and streptozocin.

Representative DNA synthesis inhibitors include (±) ametopterin (methotrexate), 3-amino-1,4-dioxide, 2,4-benzotriazine, aminopterin, cytosine bD-arabinofuranoside (Ara-C), bD-hydrochloride 5 cytosine arabinofuranoside (Ara-C), 2-fluoroadenine-9-bD-arabinofuranoside (fludarabine phosphate; F-ara-A), 5-fluoro-5'-deoxyuridine, 5-fluorouracil, ganciclovir, hydroxyurea, 6- mercaptopurine, and 6-thioguanine.

Representative DNA / RNA transcriptional regulators included actinomycin D, daunorubicin hydrochloride, 5,6-dichlorobenzimidazole 1-b-D-ribofuranoside, doxorubicin hydrochloride, homoharringtonine, and idarubicin hydrochloride.

Representative enzyme activators and inhibitors include forskolin, DL-aminoglutethimide, apicidine, Bowman-Birk inhibitor, butein, 15 (S) - (+) - camptothecin, curcumin, (-) - degueline, (-) - depudecin, doxycycline hyclate, etoposide, formestan, fostriecin sodium salt, hispidine, 2-imino-1-imidazolidinoacetic acid (cyclocreatine), oxanflatin, 4-phenylbutyric acid, roscovitine, sodium valproate, trichostatin A, tirfostine AG 34, tirfostine AG 879, urinary trypsin inhibitor fragment, valproic acid 20 (2-propylpentanoic acid), and XK469.

Representative gene regulators include 5-aza-2'-deoxycytidine, 5-azacytidine, cholecalciferol (Vitamin D3), cyglitizone, cyproterone acetate, 15-deoxy-D1214-prostaglandin J2, epitestosterone, flutamide, glycyrrizic acid ammonium salt ( glycyrrhizin), 4-hydroxy tamoxifen, mifeprisone, procainamide hydrochloride, raloxifene hydrochloride, all trans-retinal (vitamin A aldehyde), retinoic acid (vitamin A acid), 9-cis-retinoic acid, 13- cis-retinoic acid, retinoic acid p-hydroxyanilide, retinol (Vitamin A), tamoxifen, tamoxifen citrate salt, tetradecylthioacetic acid, and troglitazone.

Representative HSP-90 inhibitors include 17- (allylamino) -17-

demethoxygeldanamycin and geldanamycin.

Representative microtubule inhibitors include colchicines, dolastatin 15, nocodazole, taxanes and in particular paclitaxel, podophyllotoxin, rhizoxin, vinblastine sulfate salt, vincristine sulfate salt, and vindesine sulfate salt and vinorelbine ditartrate salt (Navelbine).

Representative agents for performing phototherapy include photoactive porphyrin 5 rings, hypericin, 5-methoxypsoralen, 8-methoxypsoralen, psoralen, and ursodeoxycholic acid.

Representative agents used as adjuncts to therapy include amifostine, 4-amino-1,8-naphthalimide, brefeldin A, cimetidine, phosphomycin disodium salt, Ieuprolide acetate salt (leuprorelin), acetate salt. of luteinizing hormone releasing hormone (LH-RH), lectin, papaverine hydrochloride, pifltrin-a, (-) - scopolamine hydrobromide, and tapsigagen.

The agents may also be anti-VEGF (vascular endothelial growth factor) agents, as such are known in the art. Several antibodies and small molecules are currently in clinical trials or have been approved to work by inhibiting VEGF, such as Avastin (Bevacizumab), SU5416, SU11248 and LADRA 43-9006. Agents may also be directed against growth factor receptors such as those from the EGF / Erb-B family such as EGF Receptor (I-1).

1 20 or Gefitinib, and Tarceva or Erlotinib), Erb-B2, receptor (Herceptin or Trastuzumab), other receptors (such as Rituximab or Rituxan / MabThera), tyrosine kinases, non-receptor tyrosine kinases, serine. / cell threonine kinases (including MAP kinases), and various other proteins whose disruption contributes to oncogenesis (such as the small / Ras and large / heterotrimeric 25 G family proteins). Several antibodies and small molecules that mark those molecules are currently in various stages of development (including approved for treatment or in clinical trials). Some of the commonly used antitumor agents in use or in clinical trials included paclitaxel, docetaxel, tamoxifen, vinorelbine, gemcitabine, cisplatin, etoposide, topotecan, anastrozole, rituximab, trastuzumab, fludaberbine, cyclophosphamide, gentlophosphamide carboplatin, interferones, and doxorubicin. The commonly used anticancer agent is paclitaxel which is used alone or in combination with other chemotherapy drugs such as: 5- FU, doxorubicin, vinorelbine, cytoxan, and cisplatin.

Combination therapy may be provided by combining two or 5 more of the above compounds.

c) Agents Treating Chrohn's Disease

There are several therapeutic methods to treat Chrohn's disease. Most people are treated first with mesalamine-containing drugs, a substance that helps control inflammation. Sulfasalazine is the commonly used of these drugs. Patients who do not benefit from this or cannot tolerate it may be put on other drugs containing mesalamine, generally known as 5-ASA agents, such as A-sacol, Dipentum, or Pentasa. Corticosteroids are often administered to control inflammation.

Immunosuppressive agents are also used to treat

Crohn's Commonly prescribed is 6-mercaptopurine and a related drug, azathioprine. Immunosuppressive agents work by blocking the immune reaction that contributes to inflammation.

Patients may be treated with combinations of these agents, for example combinations of corticosteroids and immunosuppressive drugs.

The US Food and Drug Administration has approved the drug infliximab (brand name, Remicade) for the treatment of moderate to severe Crohn's disease that does not respond to standard therapies (mesalamine, 25 corticosteroids, immunosuppressive agents) and for the treatment of fistulae. open with drain. Infiiximab is an anti-tumor necrosis factor alpha (TNF-alpha) antibody. These and other anti-TNF-alpha agents can be used to remove TNF-alpha from the colon, thus preventing inflammation without the side effects that could result if TNF-alpha was removed from blood flow outside the colon.

Antidiarrheal agents are also often administered, including diphenoxylate, loperamide, and codeine. d) Agents Treating Ulcerative Colitis

Agents that are used to treat ulcerative colitis overlap those used to treat Chrohn's disease. Examples include amino-salicylates, drugs containing 5-aminosalicylic acid (5-ASA), to help control inflammation, such as sulfasalazine, olsalazine, mesalamine, and balsalazide. They also include corticosteroids such as prednisone and hydrocortisone, and immunomodulators such as azathioprine and 6-mercapto-purine (6-MP), cytokines, interleukins, and lymphokines. Cyclosporine may be used with 6-MP or azathioprine to treat severe, active ulcerative colitis. Anti-TNF-alpha agents, thiazolidinediones or glitazones, including rosiglitazone and pioglitazone, may also be used.

e) Agents Treating Constipation / Irritable Bowel Syndrome

Constipation, such as that associated with irritable bowel syndrome, is often treated using stimulant laxatives, osmotic laxatives such as Lactulose and MiraLax, stool softeners (such as mineral oil or Colace), pasta agents (such as Metamucil or bran). wheat). Agents such as Zelnorm (also called tegaserod) can be used to treat IBS with constipation. Additionally, anticholinergic medications such as Bentyl® and Levsin® have been found to be useful in relieving intestinal spasms of IBS.

f) Protein and Peptide Drugs

Drug delivery systems may be used to orally administer proteins and peptides that could otherwise be degraded if orally administered, and which would otherwise have to be administered intramuscularly or intravenously. Examples of protein and peptide drugs useful in the present invention include:

Adrenocorticotropic hormone (ACTH) peptides including, but not limited to, human ACTH; ACTH 1-10; ACTH 1-13, human: AC-TH 1-16, human; ACTH 1-17; ACTH 1-24, human; ACTH 4-10; ACTH 4-301; ACTH 6-24; ACTH 7-38, human; ACTH 18-39, human; ACTH, rat; ACTH 12-39, rat; beta cell tropin (ACTH 22-39); biotinyl-ACTH 1-24, human; biotinyl-ACTH 7-38, human; corticostatin, human; corticostatin, rabbit; [Met (02) 4, DLys81 Phe9] ACTH 4-9, human; [Met (O) 4, DLys81 Phe9] ACTH 4-9, human; N-acetyl, ACTH 1-17, human; and ebiratide.

Adrenomedulin peptides including, but not limited to, adrenomedulin, human adrenomedulin 1-52; adrenomedulin 1-12, human 5; human adrenomedulin 13-52; human adrenomedulin 22-52; pro-adrenomedulin 45-92, human; pro-adrenomedulin 153-185, human; adrenomedulin 1-52, porcine; pro-adrenomedulin (N-20), porcine; adreno-medulin 1-50, rat; adrenomedulin 11-50, rat; and pro-N20 (proadrenomedulin N-terminal peptide 20), rat.

Alatostatin peptides including, but not limited to, alatostatin

tatine I; alatostatin II; alatostatin III; and allatostatin IV.

Amylin peptides including, but not limited to, human acetyl amylin 8-37; acetylated amylin 8-37, rat; AC 187 amylin antagonist; amylin antagonist AC253; amylin antagonist AC625; amylamine 15-8-37, human; amylin (IAPP), cat; amylin (insulinoma or amyloid islet polypeptide (IAPP)); human amylin amide; amylin 1-13 (diabetes associated peptide 1-13), human; amylin 20-29 (IAPP 20-29), human; amylin antagonist AC625; human amylin 8-37; amylin (IAPP), cat; amylin, rat; amylin 8-37, rat; biotinyl amylin, rat; and human biotinyl amylin amide.

Amyloid beta-protein fragment peptides including, but not limited to, Alzheimer's disease beta-protein 12-28 (SP17); amyloid beta protein 25-35; beta / A4-amyloid protein precursor 328-327; beta / A4 amyloid protein precursor (APP) 319 - 335; amyloid beta-protein 1-43; amyloid beta protein 1-42; amyloid beta protein 1-40; amyloid protein 10-20; amyloid beta protein 22-35; Alzheimer's disease beta protein (SP28); beta-amyloid peptide 1-42, rat; beta-amyloid peptide 1-40, rat; beta amyloid 1-11; beta amyloid 31-35; beta amyloid 32-35; beta amyloid 35-25; beta-amyloid protein precursor / 30 A4 96-110; beta-amyloid protein precursor 651-616; beta amyloid 1-38; [Gln11] -beta-protein of Alzheimer's disease; [Gln11] beta amyloid 1-40; [Gln22] beta-amyloid 6-40; non-A beta component of Alzheimer's disease (NAC) amyloid; P3, (A beta 17-40) β-amyloid peptide of Alzheimer's disease; and SAP (serum amyloid P component) 194-204.

Angiotensin peptides including, but not limited to, A-779; Ala-Pro-Gly-angiotensin II; [lle 3, Val 5] angiotensin II; angiotensin III antipeptide; angiogenin fragment 108-122; angiogenin fragment 108-123; angiotensin converting enzyme inhibitor I; angiotensin I, human; angiotensin converting enzyme I substrate; angiotensin 11-7, human; angiopeptin; human angiotensin II; angiotensin II antipeptide; human angiotensin II 1-4; human angiotensin III 3-8; human angiotensin II 4-8; human angiotensin II 5-8; angiotensin III ([Des-Asp1] -angiotensin II), human; angiotensin III inhibitor ([II7] - angiotensin III); angiotensin converting enzyme inhibitor (Neo-thunnus macropterus); [Asn1, Val5] -angiotensin I, frogfish; [Asn1, Val5, Asn9] -angiotensin I, salmon; [Asn1, Val5, Gly9] -angiotensin I, eel; [Asn1, Val5] -angiotensin I 1-7, eel, frogfish, salmon; [Asn1, Val5] - angiotensin II; human biotinyl angiotensin I; biotinyl angiotensin II, human; biotinyl-Ala-Ala-Ala-angiotensin II; [Des-Asp1] -angiotensin I, human; [p-aminophenylalanine6] -angiotensin II; renin substrate (angiotensinogen 1-13), human; preangiotensinogen 1-14 (renin substrate tetradecapeptide), human; renin substrate tetradecapeptide (angiotensinogen 1-14), porcine; [Sar1] -angiotensin II, [Sar1] -angiotensin II 1-7 amide; [Sar1, Ala8] -angiotensin II; [Sar1, lle8] -angiotensin II; [Sar1, Thr8] - angiotensin II; [Sar1, Tyr (Me) 4] -angiotensin II (Sarmesin); [Sar1, Valr, Ala8] - angiotensin II; [Sar1, 1le7] -angiotensin III; synthetic tetradeca peptide renin substrate (No. 2); [Val4] angiotensin III; [Val5] angiotensin II; [Val5] - angiotensin I, human; [Val5] angiotensin I; [Val5, Asn9] -angiotensin I, frog; and [Val5, Ser9] -angiotensin I, ave.

Antibiotic peptides including, but not limited to, Ac-SQNY; bacterenecin, bovine; CAP 37 (20-44); carbormethoxycarbonyl-DPro-DPhe-OBzl; CD36 peptide P 139-155; CD36 peptide P 93-110; cecropin A-melittin hybrid peptide [CA (1-7) M (2-9) NH2]; cecropin B, free acid; CYS CD fragment (Bzl) 84 81-92; defensin (human) HNP-2; dermaptin; immunostimulation peptide, human; Iactoferricin, bovine (BLFC); and magainin spacer.

Antigenic polypeptides, which may elicit an enhanced immune response, enhance an immune response and / or cause an effective immunizing response to disease and / or disease causing agents including, but not limited to, adenovirus; anthrax; Bordetella perussus; botulism; bovine rhinotracheitis; Branhamella catarrhalis; canine hepatitis; canine synomosis; Chlamydiae; cholera; coccidiomycosis; cowpox; cytomegalovirus; dengue fever; dengue toxoplasmosis; diphtheria; encephalitis; Enterotoxigenic Escherichia coli; Epstein Barr virus; equine encephalitis; equine infectious anemia; equine influenza; equine pneumonia; equine rhinovirus; Eseheriehia feline leukemia; flaviviruses; globulin; haemophilus influenza type b; Haemophilus influenzae] Haemophilus pertussis; Heiieobaeter pylon; hemophilus ??; hepatitis ??; hepatitis A virus; hepatitis B virus; hepatitis C virus; herpes virus; HIV; HIV-1 virus; HIV-2 virus; HTLV I; HTLV II; HTLV III; influenza? Japanese encephalitis; Klebsiellae species; Legionella pneumophila; leishmania; leprosy; lyme disease; malaria immunogen; measles; meningitis; Meningococcus; group A meningococcal polysaccharide; group C meningococcal polysaccharide; mumps; mumps virus; mycobacteria; Mycobaeterium tubereulosis, Neisseria; Neisseria gonorrhoeae \ Neisseria meningitidis; sheep blue tongue; ovine encephalitis; papilloma virus; for influenza; paramyxovirus; pertussis toxins; pestilence; pneumococcus; Pneumoeystis earinir, pneumonia; poliovirus; Proteus species; Pseudomonas aeruginosa; rage; respiratory syncytial virus; rotavirus; rubella; Salmonellae; schistosomiasis; shigellae; simian immunodeficiency virus; smallpox; Staphyloeeoeus aureus Staphyloeeoeus species; Streptoeeeus pneumoniae; Streptoeeoeus pyogens; Streptoeeoeus species; Clostridium diffieile; Clostridium species; swine influenza; tetanus; Treponema pallidum; typhoid fever; vaccinia; varicella-zoster virus; and bibrio cholerae.

Antimicrobial peptides including, but not limited to, buforin I; buforin II; cecropin A; cecropin B; cecropin P1, porcine; gaegurin 2 (Raña rugosa); gaegurin 5 (Rana rugosa); indolicidin; protegrin- (PG) -1; magainin 1; and magainin 2; and T-22 [Tyr5,12, Lys7] -poly-femusin II peptide.

Apoptosis-related peptides including, but not limited to, Alzheimer's disease beta-protein (SP28); calpain inhibitor peptide; IV capsase inhibitor; capsase-3, substrate IV; cell permeable caspase-1 inhibitor I; caspase-1 inhibitor V1; caspase-3 substrate, fluorogenic; fluorogenic caspase-1 substrate V; cell permeable caspase-3 inhibitor I; caspase-6 inhibitor ICE; [Des-Ac, biotin J-ICE inhibitor III]; IL-1B converting enzyme (ICE) inhibitor II; IL-IB converting enzyme (ICE) substrate IV; MDL 28170; and MG-132.

Atrial natriuretic peptides including, but not limited to, alpha-ANP (alpha-chANP), chicken; ananthin; ANP 1-11, rat; ANP 8-30, frog; ANP

11-30, frog; ANP-21 (fANP-21), frog; ANP-24 (fANP-24), frog; ANP-30, frog; human, canine ANP 5-28 fragment; ANP-7-23, human; human, canine 15 PNA fragment 7-28; natriuretic alpha-atrial polypeptide 1-28, human, canine; A71915, rat; natriuretic atrial factor 8-33, rat; human natriuretic atrial polypeptide 3-28; atrial natriuretic polypeptide 4-28, human, canine; atrial natriuretic polypeptide 5-27; human; natriuretic atrial peptide (ANP), eel; atriopeptin I, rat, rabbit, mouse; atriopeptin II, rat,

120 rabbit, mouse; atriopeptin III, rat, rabbit, mouse; atrial natriuretic factor (rANF), rat, auriculin A (rat ANF 126-149); auriculin B (rat ANF 126-150); beta-ANP (1-28, dimer, antiparallel); beta-rANF 17-48; biotinyl alpha-ANP 1-28, human, canine; biotinyl atrial natriuretic factor (biotinyl-rANF), rat; cardiodilatin 1-16, human; C-ANF 4-23, rat; Des- [Cys105,25 Cys121] atrial natriuretic factor 104-126, rat; [Met (O) 12] ANP 1-28, human; [Mpr7, DAIa9] ANP 7-28, amide, rat; prepro-ANF 104-116, human; prepro-ANF 26-55 (proANF 1-30), human; prepro-ANF 56-92 (proANF 31-67), human; prepro-ANF 104-123, human; [Tyr °] atropeptin I, rat, rabbit, mouse; [Tyr °] atropeptin II, rat, rabbit, mouse; [Tyr °] -pro 30 ANF 104-123, human; urodilatin (CDD / ANP 95-126); ventricular natriuretic peptide (VNP), eel; and ventricular natriuretic peptide (VNP), rainbow trout. Pouch cell peptides, but not limited to, alpha pouch cell peptide; alpha 1-9 pocket cell peptide; alpha pocket bag peptide 1-8; alpha pocket bag peptide 1-7; beta cell factor beta; and gamma bolca cell factor.

Bombesin peptides including, but not limited to, alpha-sl

casein 101-123 (bovine milk); biotinyl bombesin; bombesin 8-14; bombesin; [Leu13-psi (CH2NH) Leu14] -bombesine; [D-Phe61 Des-Met14] -bombesine 6-14 ethylamide; [DPhe12] -bombesine; [DPhe12,1114] -bombesine; [Tyr4] - bombesin; and [Tyr4, DPhe12] -bombesine.

Bone GLA (BGP) Peptides Including, But Not Limited

to, bone GLA protein; bone GLA protein 45-49; [Glu17, Gla21.24] -osteocalcin 1-49, human; myclopeptide-2 (MP-2); human osteocalcin 1-49; osteocalcin 37-49, human; and [Tyr38, Phe42'46] human 38-49 bone GLA protein.

Bradykinin peptides including, but not limited to, [Ala2,6,

des-Pro 3] -bradykinin; bradykinin; bradykinin (Bowfin. Gar); bradykinin enhancer peptides; bradykinin 1-3; bradykinin 1-5; bradykinin 1-6; bradykinin 1-7; bradykinin 2-7; bradykinin 2-9; [DPhe7] bradykinin; [Des-Arg9] bradykinin; [Des-Arg10] -Lys-bradykinin ([Des-Arg10] -calidin); [D-N-Me-Phe7] bradykinin; [Des-Arg91 Leu8] bradykinin; Lys-bradykinin (kalidine); Lys- [Des-Arg9, Leu8] -bradykinin ([Des-Arg10, Leu9] -calidine); [Lys ° -Hyp3] -bradykinin; ovoquinine; [Lys01 Ala3] - bradykinin; Met-Lys-bradykinin; K12 peptide bradykinin enhancer peptide; [(pCI) Phe5,8] bradykinin; T-kinine (Ile-Ser-bradykinin); [Thi5.8, D-Phe7] -bradykinin; [Tyr °] bradykinin; [Tyr5] bradykinin; [Tyr8] - bradykinin; and kallikrein.

Brain natriuretic peptides (BNP) including, but not limited to, canine BNP 32; BNP-like peptide, eel; BNP-32, human; BNP-45, mouse; BNP-26, porcine; BNP-32, porcine; biotinyl-BNP-32, porcine; BNP-32, rat; biotinyl-BNP-32, rat; BNP-45 (BNP 51-95, 5K cardiac natriuretic peptide), rat; and [Tyr °] -BNP 1-32, human.

C-peptides including, but not limited to, C-peptide; and [Tyr0] - C-peptide, human.

C-type natriuretic peptides (CNP) including, but not limited to, C-type natriuretic peptide, chicken; C2-type natriuretic peptide (CNP-22), porcine, rat, human; human C 5 -type natriuretic peptide (CNP-53); type C natriuretic peptide (CNP-53), porcine, rat; C-53 type natriuretic peptide (porcine, rat) 1-29 (CNP-53 1-29); prepro-CNP 1-27, rat; prepro-CNP 30-50, porcine, rat; vasonatrin peptide (VNP); and C-type [Tyr °] natriuretic peptide 22 ([Tyr °] -CNP-22).

Calcitonin peptides including, but not limited to, biotinyl

calcitonin, human; biotinyl calcitonin, rat; biotinyl calcitonin, salmon; calcitonin, chicken; calcitonin, eel; calcitonin, human; calcitonin, porcine; calcitonin, rat; calcitonin, salmon; human calcitonin 1-7; calcitonin 8-32, salmon; catacalcine (PDN-21) (C-procalcitonin); and N-proCT (amino terminal procalcitonin cleavage peptide 15), human.

Calcitonin gene-related peptides (CGRP) including, but not limited to, acetyl alpha-CGRP 19-37, human; alpha-CGRP 19-37, human; alpha-CGRP 23-37, human; biotinyl-CGRP, human; biotinyl-CGRP

II, human; biotinyl-CGRP, rat; beta-CGRP, rat; biotinyl beta-CGRP, rat; CGRP, rat; CGRP, human; calcitonin peptide adjacent to the C-terminus; CGRP 1-19, human; CGRP 20-37, human; CGRP 8-37, human; C-GRP II, human; CGRP, rat; CGRP 8-37, rat; CGRP 29-37, rat; CGRP

30-37, rat; CGRP 31-37, rat; CGRP 32-37, rat; CGRP 33-37, rat; CGRP

31-37, rat; ([Cys (Acm) 2,7] -CGRP; elcatonin; [Tyr0J-CGRP, human; [Tyr0] CGRP II, human; [Tyr °] -CGRP 28-37, rat; [Tyr °] -CGRP, rat; and [Tyr22] -

CGRP 22-37, rat.

CART peptides including, but not limited to, CART, human; CART 55-102, human; CART, rat; and CART 55-102, rat.

Casomorphine peptides including, but not limited to, human beta-casomorphine; beta-casomorphine 1-3; beta-casomorphine 1-3, amide; beta-casomorphine, bovine; beta-casomorphine 1-4, bovine; beta-casomorphine 1-5, bovine; beta-casomorphine 1-5, amide, bovine; beta-casomorphine 1-6, bovine; [DAIa] beta-casomorphine 1-3, amide, bovine; [DAIa2, Hyp4, Tyr5] beta-casomorphine 1-5 amide; [DAIa2, DPro4, Tyr5] beta-casomorphine 1-5, amide; [DAIa2, Tyr5] beta-casomorphine 1-5, amide, bovine; [DAIa2,4, Tyr5] beta-casomorphine 1-5, amide, bovine; [DAIa 2, (pCI) Phe 3] -beta-casomorphine, bovine amide; [DAIa2] beta-casomorphine 1-4, amide, bovine; [DAIa2] beta-casomorphine 1-5, bovine; [DAIa2] beta-casomorphine 1-5, amide, bovine; [DA-1a2, Met5] -beta-casomorphine 1-5, bovine; [DPro2] beta-casomorphine 1-5, amide, bovine; [DAIa2] -beta-casomorphine 1-6, bovine; [DPro2] beta-casomorphine 1-4, amide; [Des-Tyr1] -beta-casomorphine, bovine; [DAIa2,4, Tyr5] beta-casomorphine 10 1-5, amide, bovine; [DAIa 2, (pCI) Phe 3] beta-casomorphine, amide, bovine; [DA-la2] beta-casomorphine 1-4, amide, bovine; [DAIa2] beta-casomorphine 1-5, bovine; [DAIa2] beta-casomorphine 1-5, amide, bovine; [DAIa2, Met5] -betacasomorphine 1-5, bovine; [DPro2] beta-casomorphine 1-5, amide, bovine; [DA-la2] -beta-casomorphine 1-6, bovine; [DPro2] beta-casomorphine 1-4, amide; [Des-Tyr1] -beta-casomorphine, bovine; and [Val3] beta-casomorphine 1-4, bovine amide.

Chemotactic peptides including, but not limited to, defense 1 (human) HNP-I (human neutrophil peptide-1); and N-formyl-Met-Leu-Phe.

Cholecystokinin peptides (CCK) including, but not limited to

a, caerulein; cholecystokinin; cholecystokinin-pancreozimine; CCK-33, human; cholecystokinin 1-4 octapeptide (unsulfated) (CCK 26-29, unsulfated); cholecystokinin octapeptide (CCK 26-33); cholecystokinin octapeptide (unsulfated) (CCK 26-33, unsulfated); cholecystokinin heptapeptide (CCK 27-33); cholecystokinin tetrapeptide (CCK 30 - 33); CCK-33, porcine; CR 1 409, cholecystokinin antagonist; CCK flanking peptide (unsulfated); sulfated N-acetyl cholecystokinin, CCK 26-30; N-acetyl cholecystokinin, CCK 26-31, sulphated; N-acetyl cholecystokinin, CCK 26-31, unsulfated; prepro V-9-M CCK fragment 30; and proglumid.

Colony stimulating factor peptides including, but not limited to, colony stimulating factor (CSF); GM-CSF; M-CSF; and G-CSF. Corticotropin-releasing factor (CRF) peptides including, but not limited to, astressin; CRF 12-41 alpha helical; biotinyl-CRF, sheep; biotinyl CRF, human, rat; CRF, bovine; CRF, human, rat; CRF, sheep; CRF, porcine; [Cys21] -CRF, human, rat; CRF antagonist (alpha helical CRF 59-41); CRF 6-33, human, rat; [DPro5] -CRF, human, rat; [D-Phe12, N-Ie21'38J-CRF 12-41, human, rat; eosinophyllotoxic peptide; [Met (O) 21J-CRF, sheep; [NIe21jTyr32J-CRF, sheep; Prepro 125-151 CRF, human; sauvagine, frog; [Tyr10J-CRF, human, rat; [Tyr10J-CRF, sheep; [Tyr10-CRF 34-41, sheep; [Tyr ° J-urocortin; human urocortin amide; urocortin, rat; urotensin I 10 (Catostomus commersoni); urotensin II; and urothensin II (Rana ridibunda).

Cortistatin peptides including, but not limited to, cortistatin 29; cortistatin 29 (1-13); [Tyr10J-Cortistatin 29; procortistatin 28-47; and pro-cortistatin 51-81.

Cytokine peptides including, but not limited to, tumor necrosis factor alpha 15 (TNF-α); and tumor necrosis factor β (TNF-β). Interleukins, including but not limited to IL-1a, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, and IL-13. Interleukin peptides including, but not limited to, interleukin-1 beta 165-181, rat; and interleukin-8 (IL-8, CINC / gro), rat. Chemokines including but not limited to RANTES, MCP-I, 20 MIP-1a, ΜΙΡ-Ιβ.

Dermorphine peptides including, but not limited to, dermorphin and dermorphine analog 1-4.

Dynorphin peptides including, but not limited to, large dinorphin (prodynorphin 209-240), porcine; biotinyl dinorphin A (biotinyl prodynorphin 209-225); [DAIa2, DArge J-dinorphin A 1-13, porcine; [D-AIa 2 J-dinorphin A, porcine; [D-Ala2 J-dinorphin A, amide, porcine; [D-Ala2] dynorphine A 1-13, amide, porcine; [D-Ala 2] -dinorphine A 1-9, porcine; [DArg6J-dinorphin A 1-13, porcine; [DArg8J-dinorphin A 1-13, porcine; [Des-Tyr1J-dinorphin A 1-8; [D-Pro10J-dinorphin A 1-11, porcine; dinorphin A amide, porcine; dinorphin A 1-6, porcine; dinorphin A 1-7, porcine; dinorphin A 1-8, porcine; dinorphin A 1-9, porcine; dinorphin A 1-10, porcine; dinorphin A 1-10 amide, porcine; dinorphin A 1-11, porcine; dinorphin A 1-12, porcine; dinorphin A 1-13, porcine; dinorphin A 1-13 amide, porcine; DAKLI (kappa analog dynorphin A); DAKLI-biotin ([Arg11,13] -dymorphine A (1-13) -Gly-NH (CH2) 5NH-biotin); dinorphin A 2-17, porcine; dinorphin 2-17, amide, porcine; dinorphin A 2-12, porcine; dinorphin A 3-17, amide, porcine; dinorphin A 3-8, porcine; dinorphin A 3-13, porcine;

5 dinorphin A 3-17, porcine; dinorphin A 7-17, porcine; dinorphin A 8-17, porcine; dinorphin A 6-17, porcine; dinorphin A 13-17, porcine; dinorphin A (prodinorphine 209-225), porcine; dinorphin B 1-9; [MeTyr1, MeArg7, D-Leu8] -dymorphine 1-8 ethyl amide; [(nMe) Tyr1] dinorphin A I - 13, amide, porcine; [Phe7] dynorphin A 1-7, porcine; [Phe7] dynorphin A 1-7, amide, porcine; and prodinorphine 228-25610 (dinorphin B 29) (leumorphine), porcine. Endorphin peptides including, but not limited to, alpha neo-endorphin, porcine; beta neo-endorphin; Ac-beta-endorphin, camel, bovine, sheep; Ac-beta endorphin 1-27, camel, bovine, sheep; Ac-beta-endorphin, human; Ac-beta-endorphin 1-26, human; Ac-beta endorphin 1-27, human; Ac-gamma endorphin (Ac-beta lipotropin 61-1577); acetyl alpha endorphin; alpha-endorphin (beta-lipotropin 61-76); alpha neo endorphin analogue; alpha neo-endorphin 1-7; [Arg8] alpha neo-endorphin 1-8; beta-endorphin (beta-lipotropin 61-91), camel, bovine, ovine; beta-endorphin 1-27, camel, bovine, sheep; beta endorphin, equine; beta-endorphin (beta-lipotropin 61-91), human; beta endorphin (1-5) + (16-31), human; Human beta-endorphin 1-26; human beta-endorphin 1-27; human beta-endorphin 6-31; human beta-endorphin 18-31; beta endorphin, porcine; beta endorphin, rat; beta-lipotropin 1-10, porcine; beta-lipotropin 60-65; beta-lipotropin 61-64; beta lipotropin 61-69; beta-lipotropin 88-91; biotinyl beta endorphin (biotinyl beta lipotropin 61-91); human biocitin beta-endorphin; Gamma endorphin (beta-lipotropin 61-77); [DAIa2] alpha-endogenin 1-2, amide; [DAIa2] beta lipotropin 61-69; [DAIa2] gamma endorphin; [Des-Tyr1-beta-endorphin, human; [Des-Tyr1] gamma endorphin (beta-lipotropin 62-77); [Leu5] beta endorphin, camel, bovine, sheep; [Met5, Lys6] alpha-endorphin 1-6; [Met5, Lys6,7] alpha-endogenin 1-7; and [Met5, Lys6, Arg7] -alphane-endorphin 1-7.

Endothelin peptides including, but not limited to, endothelin-1 (ET-I); endothelin-1 [Biotin-Lys9]; endothelin-1 (1-15), human; endothelin-1 (1-15), amide, human; Ac-endothelin-1 (16-21), human; Ac- [DTrp16] -endothelin-1 (16-21), human; [Ala3,11] -endothelin-1; [Dprl, Asp15] - endothelin-1; [Ala2] -endothelin-3, human; [Ala18] -endothelin-1, human; [Asn18] -endothelin-1, human; [Res-701-1] - endothelin B receptor antagonist; Suc- [Glu9, Ala11,15] -endothelin-1 (8-21), IRL-1620; endothelin-C-terminal hexapeptide; Large [D-Val] -endothelin-1 (16-38), human; endothelin-2 (ET-2), human, canine; endothelin-3 (ET-3), human, rat, porcine, rabbit; biotinyl endothelin-3 (biotinyl ET-3); preproendothelin-1 (94-109), porcine; BQ-518; BQ-610; BQ-788; endothelium-dependent relaxation antagonist; FR139317; IRL-1038; JKC-301; JKC-302; PD-145065; PD 142893; sarafotoxin S6a (atractaspis engaddensis); sarafotoxin S6b (α-tractaspis engaddensis); sarafotoxin S6c (atractaspis engaddensis; [Lys4] - sarafotoxin S6c; sarafotoxin S6d; human large endothelin-1; biotinyl-large human endothelin-1; porcine endothelin-1 large; -3 large (22-41), amide, human; large endothelin-1 (22-39), rat; large endothelin-1 (1-39), bovine; large endothelin-1 (22-39), bovine ; large endothelin-1 (19-38), human; large endothelin-1 (22-38), human; large endothelin-2, human; large endothelin-2 (22-37), human; large na-3, human; large endothelin-1, porcine; large endothelin-1 20 (22-39) (prepro-endothelin-1 (74-91)); large endothelin-1, rat; large endothelin-2 (1 -38), human; large endothelin-2 (22-38), human; large endothelin-3, rat; human biotinyl-large endothelin-1, and [Tyr123] -proprendothelin (110-130), amide, human

ETa receptor antagonist peptides including, but not limited to, [BQ-123]; [BE18257B]; [BE-18257A] / [W-7338A]; [BQ-485]; FR139317; PD-151242; and TTA-386.

ETb receptor antagonist peptides including, but not limited to, [BQ-3020]; [RES-701-3]; and [IRL-1720],

Encephalin peptides including, but not limited to, adrenorphine, free acid; amidorfin (proencephaline A (104-129) -NH 2), bovine; BAM-12P (bovine adrenal medullary dodecapeptide); BAM-22P (bovine adrenal medulla docosapeptide); benzoyl-Phe-Ala-Arg; encephalin; [D-Ala1, D-1a5] -encephaline; [D-Ala2, D-Met5] -encephaline; [DAIa2] -Leu enkephalin, amide; [DAIa2, Leu5, Arg6] -encephaline; [Des-Tyr1, DPen25] -encephaline; [Des-Tyr1, DPen2, Pen5] -encephaline; [Des-Tyr1] -Leu enkephalin; [D-Pen2,5] encephalin; [DPen2, Pen5] -encephaline; encephalinase substrate; [D-Pen2, pCI-Phe4, D-Pen5] -encephaline; Leu-enkephalin; Leu-enkephalin, amide; biotinyl-Leu-enkephalin; [D-Ala2] -Leu enkephalin; [D-Ser1 -Leu-enkephalin-Thr (delta receptor peptide) (DSLET); [D-Thr 2 -Leu enkephalin-Thr (DTLET); [Lys6] - Leu-enkephalin; [Met5, Arg6] -encephaline; [Met5, Arg6] -encephaline-Arg; [Met51Arg6, Phe7] -encephaline, amide; Methacephalin; biotinyl Met-enkephalin; [D-Ala2] -Met-enkephalin; [D-Ala2] -Met-encephalin, amide; Met-encephalin-Arg-Phe; Methacephalin, amide; [Ala 2] -Met-enkephalin, amide; [DMet2, Pro5] - enkephalin, amide; [DTrp2] -Met-enkephalin, amide, methofinamide (adrenorphine); peptide B, bovine; 3200 Dalton adrenal E peptide, bovine; F peptide, bovine; preproencephaline B 186-204, human; spinorphin, bovine; and thiorphan (D, L, 3-mercapto-2-benzylpropanoyl glycine).

Ephrin B, its analogues and antagonists.

Fibronectin peptides including, but not limited to, platelet factor-4 (58-70), human; echistatin (Eehis carinatus); conserved region of selectin Ε, P, L; fibronectin analog; fibroectin binding protein; fibrinopeptide A, human; [Tyr °] fibrinopeptide A, human; fibrinopeptide B, human; [Glu1] fibrinopeptide B, human; [Tyr15] - fibrinopeptide B, human; 24-42 fibrinogen beta chain fragment; fibrinogen binding inhibitor peptide; fibronectin-related peptide (collagen binding fragment); fibrinolysis inhibition factor; FN-C / H-1 (fibronectin heparin binding fragment); FN-C / H-V (fibronectin heparin binding fragment); heparin binding peptide; laminin penta peptide, amide; Leu-Asp-Val-NH 2 (LDV-NH 2), human, bovine, rat, chicken; necrofibrin, human; necrofibrin, rat; and platelet membrane glycoprotein peptide IIB 296-306. Galanin peptides including, but not limited to, galanin,

human; galanine 1-19, human; preprogalanine 1-30, human; preprogallin 65-88, human; preprogalanine 89-123, human; galanin, porcine; galanine 1-16, porcine, rat; galanin, rat; biotinyl galanine, rat; prepro-Ianine 28-67, rat; 1-13-bradykinin 2-9 galanine, amide; M40, galanine 1- 13 -Pro-Pro- (AIa-Leu) 2-Ala-amide; C7, 1-13-aspartan-amide galanine; GMAP 1-41, amide; GMAP 16-41, amide; GMAP 25-41, amide; galanthoid; and entero-cassinine.

Gastrin peptides including, but not limited to, gastrin, chicken; human gastric inhibitor peptide (GIP); gastrin I, human; human biotinyl gastrin I; large gastrin-1, human; gastrin release peptide, human; human gastrin release peptide 1-16; gastric inhibitor polypeptide (GIP), porcine; gastrin release peptide, porcine; biotinyl gastrin release peptide, porcine; human porcine 14-27 gastrin release peptide; small gastrin, rat; pentagastrin; gastric inhibitor peptide 1-30, porcine; gastric inhibitor peptide 1-30, amide, porcine; [Tyr °] -peptide 23-42 gastric inhibitor, human; and gastric inhibitor peptide, rat.

Glucagon peptides including, but not limited to, [Des-His1, Glu9] -glucagon, extendin-4, glucagon, human; biotinyl glucagon, human; glucagon 19-29, human; glucagon 22-29, human; Des-His1- [Glu9] glucagon, amide; glucagon 1-like peptide, amide (preproglucagon 72-107, amide); glucagon 1-like peptide (preproglucagon 72-108), human; glucagon 1 (7-36) -like peptide (preproglucagon 78-107, amide); glucagon II-like peptide, rat; glucagon-1 (7-36) biotinyl-like peptide (biotinyl-preproglucagon 78-107, amide); glucagon 2-like peptide (preproglucagon 126-159), human; oxyntomodulin / glucagon 37; and valosine (VQY peptide), porcine.

Gn-RH-associated peptides (GAP) including, but not limited to, human 25-53-associated Gn-RH peptide; Gn-RH 1-24 associated peptide, human; human Gn-RH 1-13-associated peptide; Gn-RH 1-13 associated peptide, rat; human follicular gonadotropin release peptide; [Tyr °] -interval ([Tyr °] -Gn-RH Precursor Peptide 14-69), human; and propiomelanocortin precursor (POMC) 27-52, porcine.

Growth factor peptides including, but not limited to, cell growth factors; epidermal growth factors; tumor growth factor; TGF-alpha, human; TGF-alpha, from other mammalian TGF-beta species; alpha-TGF 34-43; Human EGF (epidermal growth factor); acidic fibroblast growth factor; basic fibroblast growth factor; basic fibroblast growth factor 13-18; basic fibroblast growth factor 120-125; brain-derived acidic fibroblast growth factor 1-11; brain-derived basic fibroblast growth factor 1-24; brain-derived acidic fibroblast growth factor 102-111; [Cys (Acm20,31)] - epidermal growth factor 20-31; epidermal growth factor receptor peptide 985-996; insulin-like growth factor (IGF) -I, chicken; IGF-I, rat; Human IGF-II; Des (1-3) IGF-I, human; IGF-I R3, human; IGF-I R3, human; Long IGF-I R3, human; adjuvant peptide analog; anorectic peptide; Des (1-6) IGF-II, human; IGF-II R6, human; IGF-I analog; IGF I (24-41); IGF I (57-70); IGF I (30-41); IGF II; IGF II (33-40); [Tyr °] -IGF II (33 - 40); liver cell growth factor; midcin; midcin 60-121, human; N-acetyl, alpha-TGF 34-43, methyl ester, rat; nerve development factor (NGF), mouse; platelet-derived growth factor; platelet-derived growth factor antagonist; Ligands for Erb-B family receptors.

Growth hormone peptides including, but not limited to, human growth hormone (hGH); growth hormone 1-43, human; growth hormone 6-13, human; growth hormone release factor, human; growth hormone release factor, bovine; growth hormone release factor, porcine; growth hormone release factor 1-29, amide, rat; hormone pro-release growth factor, human; biotinyl growth hormone release factor, human; growth hormone release factor 1-29, amide, human; [D-Ala2] - growth hormone release factor 1-29, amide, human; [N-Ac-Tyr1, D-Arg2] -GRF 1-29, amide; [His1, Nle27] growth hormone releasing factor 1-32, amide; growth hormone release factor 1-37, human; growth hormone release factor 1-40, human; growth hormone release factor 1-40, amide, human; growth hormone release factor 30-44, amide, human; growth hormone release factor, mouse; growth hormone release factor, sheep; growth hormone release factor, rat; biotinyl growth hormone release factor, rat; GHRP-6 ([His1, Lys6] - GHRP); hexarrelin (growth hormone releasing hexapeptide); and [D-Lys3] -GHRP-6.

GTP binding proteins and fragment peptides thereof including, but not limited to, [Arg8] -GTP binding protein fragment, alpha Gs; GTP-binding protein fragments of the beta G family; GTP binding protein fragments of the G gamma family; GTP binding protein fragment, Galfa; GTP binding protein fragments, Go alpha a and b; GTP binding protein fragment, alpha Gs; and GTP binding protein fragments, G1 alpha, G1 alpha, G13 alpha; GTP binding protein fragment, Golf alfa; GTP binding protein fragment, alpha Gz; GTP binding protein fragment, alpha Gq.

Guaniline peptides including, but not limited to, guaniline, human; guaniline, rat; and uroguaniline.

Inhibin peptides including, but not limited to, inhibin, bovine; inhibit, human alpha-subunit 1-32; [Tyr °] -inhibin, alpha-subunit 1-32, human; human seminal plasma inhibin-like peptide; [Tyr °] -peptide similar to seminal plasma inhibin, human; inhibin, alpha subunit 1-32, porcine; and [Tyr °] -inhibin, alpha-subunit 1-32, porcine.

Interferon peptides including, but not limited to, interferon alpha species (e.g., alfal, alpha2, alpha2a, alpha2b, alpha2c, alpha2d, alpha3, alpha4, alpha4a, alpha4b, alpha5, alpha ?, alpha74, alpha76 , alphaA, alfaB, alphaC, alphaCI, alfaD, alfaE, alfaF, alfaG, alfaG, alfaH, alfal, alfaJI, alfaJ2, alfaK, alfaL); interferon beta species (e.g., betaa); gamma interferon species (e.g., gamma a, gamma b); interferon epsil; interferon tau; omega interferon or any omega interferon analog. Several interferon gamma analogs are described in Pechenov et al. "Methods for preparation of recombinant cytokine proteins V. mutant analogues of human interferon-gamma with higher stability and activity" Protein Expr. Purif. 24: 173-180 (2002) are incorporated herein by reference in their entirety for teachings directed to the preparation and testing of interferon analogs.

Insulin peptides including, but not limited to, human insulin; insulin, porcine; Human IGF-II; insulin-like growth factor II (69-84); proinsulin-like growth factor II (68-102), human; proinsulin-like growth factor II (105-128), human; [AspB28] -insulin, human; [LysB28] -insulin, human; [Leu B28] - insulin, human; [Val B28] -insulin, human; [Ala B28] -insulin, human; [Asp828, ProB29] -insulin, human; [Lys628, ProB29] -insulin, human; [Leu828, ProB29] -insulin, human; [ValB28, ProB29] -insulin, human; [Ala828, Pro629] - insulin, human; [γ3lyA21] -insulin, human; [Gl / 21, GlnB3] -insulin, human; [AlaA21] -insulin, human; [AlaA21, GlnB3] -insulin, human; [GlnB3] -insulin, human; [GlnB30] -insulin, human; [Gl / 21 'GluB30] -insulin, human; [Gl / 21 'GlnB3] insulin, human; [Gln63, GluB30] -insulin, human; human B22-B30 insulin; human B23-B30 insulin; human B25-B30 insulin; human B26-B30 insulin; human B27-B30 insulin; human B29-B30 insulin; the human insulin A chain, and the human insulin B chain.

Iaminin peptides including, but not limited to, laminin; alpha 1 (1) -CB 3 435-438, rat; and laminin binding inhibitor.

Ieptin peptides including, but not limited to, human Ieptin 93-105; Ieptin 22-56, rat; Tyr-Ieptin 26-39, human; and Ieptin 116-130, amide, mouse.

Ieucoquinin peptides including, but not limited to, Ieucomosuppressin (LMS); Eukopyroquinine (LPK); Ieukokinin I; Euucoquinin II; Eukokinin III; Ieukokinin IV; Ieukokinin VI; Eukoquinin VII; and eukokinin VIII. Iuteinizing hormone releasing hormone peptides including, but not limited to, antide; Gn-RH II, chicken; luteinizing hormone releasing hormone (LH-RH) (GnRH); biotinyl-LH-RH; cetrorelix (D-20761); [D-AIa6J-LH-RH; [Gln8] -LH-RH (chicken LH-RH); [DLeu6, Val7] LH-RH 1-9, ethyl amide; [D-Lys6J-LH-RH; [D-Phe21 Pro3, D-Phe6] -LH-RH; [DPhe 2, DAIa 6] -LH-RH; [Des-Gly10] -LH-RH, ethyl amide; [D-Ala61 Des-Gly10] -LH-RH, ethyl amide; [DTrp6] -LH-RH, ethyl amide; [D-Trp6, Des-GIy10J-LH-RH, ethyl amide (deslorelin); [DSer (But) 6, Des-GIy10J-LH-RH, ethyl amide; ethyl amide; leuprolide; LH-RH 4-10; LH-RH 7-10; LH-RH, free acid; LH-RH, lamprey; LH-RH, salmon; [Lys8J-LH-RH; [Trp71Leu8J LH-RH, free acid; and [(t-Bu) DSer6, (Aza) GIy10J-LH-RH.

Mastoparan peptides including, but not limited to, mastoparan; mas7; mas8; mas17; and mastoparan X.

Mast cell degranulating peptides including, but not limited to, mast cell degranulating peptide HR-1; and mast cell degranulating peptide H-2.

Melanocyte hormone stimulating peptides (MSH) including, but not limited to, [Ac-Cys4lDPhe7lCys10J alpha-MSH 4-13, amide; alpha-melanocyte stimulating hormone; alpha-MSH, free acid; beta-MSH, porcine; biotinyl alpha-melanocyte stimulating hormone; biotinyl- [Nle4, D-Phe7] alpha-melanocyte stimulating hormone; [Des-Acetyl] alpha-MSH; [DPhe7J-alpha-MSH, amide; gamma-1-MSH, amide; [Lys °] gamma-1-MSH, amide; MSH release inhibiting factor, amide; [Nle 4] alpha-MSH, amide; [Nle4, D-Phe7J-alpha-MSH; N-Acetyl, [NIe41DPhe7J alpha-MSH 4-10, amide; beta-MSH, human; and gamma-MSH.

Morphiceptin peptides including, but not limited to, morphiceptin (beta-casomorphine 1-4 amide); [D-Pro] morphorptine; and [N-MePhe31D-Pro4] morphorptine.

Motilin peptides including, but not limited to, canine motilin; motilin, porcine; biotinyl motilin, porcine; and [Leu13] -motiline, porcine.

Neuropeptides including, but not limited to, Ac-Asp-GIu; achatin 1 cardioexcitatory peptide (ACEP-I) (Achatina fulica); adipokinetic hormone (ΑΚΗ) (Locust); adipokinetic hormone (Heliothis zea and Manducauca); alitesin; adipokinetic hormone of Tabanus atratus (Taa- AKH); adipokinetic hormone Il (Locusta migratory); adipokinetic hormone Il (Schistocera gregaria); adipokinetic hormone III (AKH-3); adipokinetic hormone G (AKH-G) (Gryllus bimaculatus); alatotropin (A) (Manduca sexta); alatotropin 6-13 (Manduca Sixth); APGW amide (Lymnaea stagnalys); buccal; cerebellin; [Des-Ser1] -cerebellin; corazonin (American Cocochroach Periplaneta americana); cardioactive crustacean peptide (CCAP); crustacean erythophore; DF2 (Procambarus clarkii); human diazepam binding inhibitor fragment; diazepam binding inhibitor (ODN) fragment; eledoisin-related peptide; FMRF amide (mollusc cardioexcitatory neuropeptide); Gly-Pro-Glu (GPE), human; granuliberrin R; head activator neuropeptide; [His7] -corazonin; hypertrea-lossemic factor of stickman; hypothalosemic hormone of Tabanus atratus (Taa-HoTH); isoguvacin hydrochloride; bicuculline methiodide; piperidine-4-sulfonic acid; proopiomelanocortin junction peptide (POMC), bovine; junction peptide, rat; KSAYMRF (P. redivivus) amide; cassinine; kinetetine; levitide; litorin; LUQ 81-91 (Aplysia califomica)] LUQ 83-91 (A-plysia califomica)] myoactive peptide I (Periplanetin CC-I) (Neurohormone D); myoactive peptide II (Periplanetin CC-2); myomodulin; neuron specific peptide; neuron specific enolase 404-443, rat; FF neuropeptide; porcine K1 neuropeptide; NEI neuropeptide (prepro-MCH 131-143), rat; NGE neuropeptide (prepro-MCH 110-128), rat; NFI (Procambarus clarkii)] PBAN-I (Bombyx mod); Hez-PBAN (Heliothis zea); SCPB (cardioactive aplisia peptide); secretoneurin, rat; uperolein; ureaquikquinine I; uquistakiquinin II; xenopsin I-related peptide; xenopsin II-related peptide; pedal peptide (Pep), aplisia; F1 peptide, lobster; phyllomedusine; mastoparan of polysts; proctoline; rana tensin; Ro I (Lubber Grasshopper, Romalea microptera); Ro Il (Lubber Grasshopper, Romalea microptera); SALMF Amide 1 (SI); SALMF amide 2 (S2); and SCPA.

Neuropeptide Y (NPY) peptides including, but not limited to, [Leu31, Pro34] -neuropeptide Y, human; neuropeptide F (Moniezia expansa); NPY antagonist B1BP3226; Bis (31/31 ') {[Cys31, Trp32, Nva34] NPY 31-36}; neuropeptide Y, human, rat; human neuropeptide Y 1-24 amide; biotinyl neuropeptide Y; [D-Tyr27,36, D-Thr32 J-NPY 27-36; Des 10-17 (cycle 7-21) [Cys7'21, Pro34] -NPY; C2-NPY; [Leu31, Pro34] neuropeptide Y, human; neuropeptide Y, free acid, human; neuropeptide Y, free acid, porcine; prepro NPY 68-97, human; N-acetyl- [Leu28, Leu31] NPY 24-36; neuropeptide Y, porcine; [D-Trp32] -neuropeptide Y, porcine; [D-Trp32] NPY 1-36, human; [Leu17jDTrp32] neuropeptide Y, human; [Leu31, Pro34] -NPY, porcine; NPY 2-36, porcine; NPY 3-36, human; NPY 3-36, porcine; NPY 13-36, human; NPY 13-36, porcine; NPY 16-36, porcine; NPY 18-36, porcine; NPY 20-36; NFY 22-36; NPY 26-36; [Pro34] -NPY 1-36, human; [Pro34] - neuropeptide Y, porcine; PYX-I; PYX-2; T4- [NPY (33-36)] 4; and Tyr (OMe) 21] - human neuropeptide Y. Neurotropic factor peptides including, but not limited to,

glial-derived neurotropic factor (GDNF); brain derived neurotropic factor (BDNF); and ciliary neurotropic factor (CNTF).

Notch receptor ligands including but not limited to proteins similar to Delta 1, Delta 2, Delta 3, Delta 4, Jagged-1 and Jagged-2, and fragments thereof.

Orexine peptides including, but not limited to, orexin A; orexin B, human; orexin B, mouse, mouse.

Opioid peptides including, but not limited to, 90-95 alpha casein fragment; BAM - 18P; casomoquinine L; casoxin D; crystalline; DALDA; dermencephaline (deltorphin) (Phylomedusa sauvagei); [D-Ala2] - deltorphin I; [D-Ala2] deltorphin II; endomorphine-1; endomorphine-2; chiotorphine; [DArg2] -kiotorfin; morphine tolerance peptide; morphine modulating peptide, C-terminal fragment; morphine modulating neuropeptide (A-18-F-NH2); nociceptin [orphan orphan CF] (ORLI agonist); TIPP; Tyr-MIF-I; Tyr-W-MIF-I; valorphin; Human LW-hemorphine-6; Leu-valorphin-Arg; and Z-pro-D-Leu.

Oxytocin peptides including, but not limited to, [Asub] - oxytocin; oxytocin; biotinyl oxytocin; [Thr4, Gly7] oxytocin; and tocinic acid ([lle3] -pressinic acid).

PACAP (Pituitary Adenylating Cyclase Activation Peptide) peptides including, but not limited to, PACAP 1-27, human, sheep, rat; PACAP (1-27) -Gly-Lys-Arg-NH2, human; [Des-GIn16J-PACAP 6-27, human, sheep, rat; PACAP38, frog; PACAP27-NH2, human, sheep, rat; biotinyl-PACAP27-NH2, human, sheep, rat; PACAP 6-27, human, sheep, rat; PACAP38, human, sheep, rat; biotinyl-PACAP38, human, sheep, rat; PA-CAP 6-38, human, sheep, rat; PACAP27-NH2, human, sheep, rat; biotinyl-PACAP27-NH2, human, sheep, rat; PACAP 6-27, human, sheep, rat; PACAP38, human, sheep, rat; biotinyl-PACAP38, human, sheep, rat; PA-CAP 6-38, human, sheep, rat; PACAP38 16-38, human, sheep, rat; PA-CAP38 31-38, human, sheep, rat; PACAP38 31-38, human, sheep, rat; PACAP-related peptide (PRP), human; and PACAP-related peptide (PRP), rat.

Pancreastatin peptides including, but not limited to, bovine chromostatin; pancreastatin (hPST-52) (chromogranin A 250-301, amide); pancreastatin 24-52 (hPST-29), human; chromogranin UNS 286-301, amide, human; pancreastatin, porcine; biotinyl pancreastatin, porcine; [Nle 8] pancreastatin, porcine; [Tyr0, Nle8] -pancreastatin, porcine; [Tyr0] - pancreastatin, porcine; parastatin 1-19 (chromogranin A 347-365), porcine; pancreastatin (chromogranin A 264-314-amide, rat; biotinyl pancreastatin (biotinyl-chromogranin A 264-314-amide; [Tyr °] -pancreastatin, rat; pancreastatin 26-51, rat; and pancreastatin 33-49, porcine.

Pancreatic polypeptides including, but not limited to, avian pancreatic polypeptide; human pancreatic polypeptide; human C-fragment pancreatic polypeptide acid; human C1-fragment pancreatic polypeptide amide; pancreatic polypeptide (temporary Rana); pancreatic polypeptide, rat; and pancreatic polypeptide, salmon.

Parathyroid hormone peptides including, but not limited to, human [Asp76] parathyroid hormone 39-84; [Asp76] parathyroid hormone 53-84, human; hormone 64-84, human; [Asn, Leu18] -parathyroid hormone 1-34, human; [Cys5> 28] -parathyroid hormone 1-34, human; hypercalcemia malignancy factor 1-40; [Leu] parathyroid hormone 1-34, human; [Lys (biotinyl) 13, Nle8'18, Tyr34] parathyroid hormone 1-34 amide; [Nle8'18, Tyr34] -parathyroid hormone 1-34 amide; [Nle8'18, T-yr34] parathyroid hormone 3-34 amide, bovine; [Nle8'18, Tyr34] parathyroid hormone 1-34, human; [Nle8'18, Tyr34] - parathyroid hormone 1-34 amide, human; [Nle8'18, Tyr34] -parathyroid hormone 3-34 amide, human; [Nle8'18, Tyr34] -parathyroid hormone 7-34 amide, bovine; [Nle8'21, Tyr34] - parathyroid hormone 1-34 amide, rat; parathyroid hormone 44-68, human; parathyroid hormone 1-34, bovine; parathyroid hormone 3-34, bovine; parathyroid hormone 1-31 amide, human; parathyroid hormone 1-34, human; parathyroid hormone 13-34, human; parathyroid hormone 1-34, rat; parathyroid hormone 1-38, human; parathyroid hormone 1-44, human; parathyroid hormone 28-48, human; parathyroid hormone 39-68, human; parathyroid hormone 39-84, human; parathyroid hormone 53-84, human; parathyroid hormone 69-84, human; parathyroid hormone 70-84, human; [Pro34] -peptide YY (PYY), human; [Tyr0] -calcemia malignancy factor 1-40; [Tyr °] parathyroid hormone 1-44, human; [Tyr0] parathyroid hormone 1-34, human; [T-yr1] -parathyroid hormone 1-34, human; [Parathyroid thyroid hormone 27-48, human; [Tyr34] parathyroid hormone 7-34 amide, bovine; [Tyr43] - parathyroid hormone 43-68, human; [Tyr52, Asn76] parathyroid hormone 52-84, human; and [Tyr63] parathyroid hormone 63-84, human.

Parathyroid hormone (PTH) -related peptides including, but not limited to, PTHrP (PTHrP 1-36 [Tyr36] -amide), chicken; hHCF- (1-34) -NH2 (humoral hypercalcemic factor), human; human PTH-related protein 1-34; biotinyl-PTH 1-34-related protein, human; PTH-related [Tyr °] protein 1-34, human; Human PTH-related protein [Tyr34] -amide 1-34; human PTH-related protein 1-37; ] - PTH-related protein amide 7 - 34, human; 38-64 PTH-related protein human amide; PTH-related protein amide] 67 - 86, human; PTH-related protein 107-111, human, rat, mouse; PTH-related protein free acid 107-111; PTH-related protein 107-138, human; and human PTH-related protein 109-111.

T-peptide peptides including, but not limited to, T-peptide; [D-Ala1] -peptide T; and T-peptide [D-Ala1] -amide.

Prolactin-releasing peptides including, but not limited to, human prolactin-releasing peptide 31; human prolactin release peptide 20; prolactin release peptide 31, rat; prolactin release peptide 20, rat; prolactin release peptide 31, bovine; and bovine prolactin release peptide 20.

YY peptide peptides (PYY) including, but not limited to, PYY, human; PYY 3-36, human; biotinyl-PYY, human; PYY, porcine, rat; and [Leu31, Pro34] -PYY, human.

Renin substrate peptides including, but not limited to, acetyl, angiotensinogen 1-14, human; angiotensinogen 1-14, porcine; renin substrate tetradecapeptide, rat; Renin substrate [Cys8] -tetradecapeptide, rat; Renin substrate [Leu8] -tetradecapeptide, rat; and renin substrate [Val8] -tetradecapeptide, rat.

Secretin peptides including, but not limited to, canine secretin; secretin, chicken; secretin, human; biotinyl secretin, human; secretin, porcine; and secretin, rat.

Somatostatin Peptides (GIF) including, but not limited to, BIM-23027; biotinyl somatostatin; biotinylated cortystatin 17, human; cortystatin 14, rat; cortistatin 17, human; [Tyr °] -cortistatin 17, human; cortystatin 29, rat; [D-Trp8] -somatostatin; [DTrp8, DCys14] -somatostatin; [D-Trp8, Tyr11] -somatostatin; [D-Trp11] -somatostatin; NTB (Naltriben); [Nle8] - somatostatin 1-28; octreotide (SMS 201-995); prosomatostatin 1-32, porcine; [Tyr °] -somatostatin; [Tyr1] -somatostatin; [Tyr1] -somatostatin 28 (1-14); [Tyr11] -somatostatin; [Tyr0, D-Trp8] -somatostatin; somatostatin; somatostatin antagonist; somatostatin-25; somatostatin-28; somatostatin 28 (1-12); biotinyl somatostatin-28; [Tyr °] -somatostatin-28; [Leu8, D-Trp22, Tyr25Homatostatin; S; biotinyl- [Leu8, D-Trp22, Ty2-somatostatin-28; somatostatin-28 (1-14); and somatostatin analog, RC-160.

Substance P peptides including, but not limited to, G protein antagonist-2; Ac- [Arg6, Sar91 Met (02) 11] -substance P 6-11; [Arg3] - substance P; Trp-3,5-bis (trifluoromethyl) benzyl Ac-ester; Ac- [Arg6, Sar9, Met (02) 11] -substance P 6-11; [D-Al4] -substance P 4-11; [Tyr6, D-Phe71 D-His9] -substance P 6-11 (sendida); biotinyl-substance P; biotinyl-NTE [Arg3] - substance P; [Tyr8] -substance P; [Sar9, Met (02) 11] -substance P; [D-Pro21 D-Trp7,9] -substance P; [D-Pro4,0-Trp7,9] -substance P 4-11; substance P 4-11; [DTrp2'7.9] -substance P; [(Dehydro) Pro2.4, Pro9] -substance P; [Dehydro-Pro4] - substance P 4-11; [Glp5, (Me) Phe8, Sar9] -substance P 5-11; [GIp51Sar9] - substance P 5-11; [Glp5] -substance P 5-11; hepta substance P (substance P 5-11); hexa-substance P (substance P 6-11); [MePhe8, Sar9] -substance P; [Nle11] -substance P; Octa-substance P (substance P 4-11); [pG | u1] -hexa- substance P ([pGlu6] -substance P 6-1 1); [pGIu6, D-Pro9] -substance P 6-11; [(pNO2) Phe7 Nle11] -substance P; penta-substance P (substance P 7-11); [Pro9] -substance P; GR73632, substance P 7-11; [Sar4] -substance P 4-11; [Sar9] -substance P; septide ([pGIu6, Pro9] -substance P 6-11); astonished I; astonished II; substance P; substance P1 cod; substance P1 trout; substance P antagonist; substance P-Gly-Lys-Arg; substance P 1-4; Substance P 1-6; substance P 1-7; substance P 1-9; deca substance P (substance P 2-11); ninth substance P (substance P 3-11); tetrapeptide of substance P (substance P 8-11); tripeptide of substance P (substance P 9-11); substance P, free acid; methyl ester of substance P; and [Tyr8lNIe11] substance P.

Tachykinin peptides including, but not limited to, [Ala51

beta-Ala8] neuroquinine A 4-10; eledoisin; Iocustataquikinin I (Lom-TK-I) (Migratory Locust); Iocustataquinine II (Lom-TK-II) (Migratory Locust); neuroquinine A 4-10; neurokinin A (neuromedine L1 substance K); neurokinine A, cod and trout; biotinyl neuroquinine A (biotinyl neuromedine L1 biotinyl substance K); [Tyr °] -neuroquinine A; [Tyr6] -substance K; FR64349; [Lys31 Gly8- (R) -gam-lactam-Leu9] -neuroquinine A 3-10; GR83074; GR87389; GR94800; [Beta-Ala8] -neuroquinine A 4-10; [Nle10] -neuroquinine A 4-10; [Trp7, beta-Ala8] -neuroquinine A 4-10; neurokinin B (neuromedin K); biotinyl neuroquinine B (biotinyl neuromedine K); [MePhe7] -ieuroquinine B; [Pro7] neurokinin B; [Tyr] neurokinin B; neuromedine B, porcine; bio-tinyl neuromedine B, porcine; neuromedine B-30, porcine; neuromedine B-32, porcine; neuromedine B receptor antagonist; neuromedine C, porcine; porcine N1 neuromedine; neuromedine (U-8), porcine; neuromedine (U-25), porcine; neuromedine U, rat; gamma neuropeptide (gamma-preprotequinine 72-92); PG-KII; phylolitorin; [Leu8] phyllolitorin (Phyllomedusa sauvagei); fisalemine; fisalemine 1-11; sciliorinin II, amide, squal; senkide, selective neurokinin B receptor peptide; [Sei] J-neuromedine C; beta-preprotaquinine 69-91, human; beta preprotaquinine 111-129, human; tachyplesin I; xenopsin; and xenopsin 25 (xenin 25), human.

Thyrotropin-releasing hormone (HRT) peptides including, but not limited to, biotinyl-thyrotropin-releasing hormone; [Glu1] - TRH; His-prodicopiperazine; [3-Me-His2] -TRH; pGlu-GIn-proamide; pGlu-His; [Phe2] -TRH; prepro TRH 53-74; prepro TRH 83-106; prepro-TRH 160-169 (Ps4, TRH- enhancer peptide); prepro-TRH 178-199; thyrotropin release hormone (HRT); TRH, free acid; TRH-SH Pro; and TRH precursor peptide.

Toxin peptides including, but not limited to, omega-agatoxin TK; agelenin (spider, Agelena opulent); apamine (honey bee, Apis mellifera); calcicudin (CaC) (green mamba, Dedroaspis angusti- ceps); calciseptin (black mamba, Dendroaspis polilepis polilepis); caribdothoxine (ChTX) (scorpion, Leiurus quinquestriatus var. hebraeus); chlorotoxin; conotoxin G1 (marine snail, Conus geographus); GS conotoxin (marine snail, Conus geographus); conotoxin M1 (Marine Magus conus); alpha-conotoxin El, Conus ermineus; alpha conotoxin SIA; alpha-conotoxin Im1; alpha-conotoxin Sl (coniferous snail, Conus striatus; GIIIB microconotoxin (marine snail, Conus geographus); omega-conotoxin GVIA (sea snail, Conus geographus); omega-conotoxin MVIIA (Conus magus); omega-conotoxin MVIIC (Conus magus); SVIB omega-conotoxin (conifer snail, Conus striatus); endotoxin inhibitor; geografutoxin I (GTX-I) (μ-Conotoxin GUIA); iberiotoxin (IbTX) (scorpion, Buthus tamulus); na 1-37; caliotoxin (scorpion, Androct-onus mauretanicus mauretanicus); mast cell degranulation peptide (MCD-peptide, peptide 401); margatoxin (MgTX) (scorpion, Centruriodes margaritatus); NSTX-3 neurotoxin (pupua nova guinea spider) , Nephilia maculata); PLTX-II (spider, Plectreurys tristes); scilatoxin (leiurotoxin I); and stichodactyl toxin (ShK); diphtheria toxin; ricin A; Pseudomonas aeruginosa A exotoxin A.

Immunotoxins consist of toxins covalently linked to an antibody that acts as a direction correction system, specifically targeting the toxin to those cells to be deleted by means of an antibody (polyclonal or monoclonal) directed against a molecule, or a group. of molecules, loaded on the surface of labeled cells. Toxins including but not limited to those cited above may be used for this purpose. The invention described in this patent application may be used for colon immunotoxin release. In some cases, the antibody may be replaced by a small molecule that similarly acts to target the toxin to a chosen group of cells.

Vasoactive intestinal peptides (VIP / PHI) including, but not limited to, VIP, human, porcine, rat, sheep; VIP-Gly-Lys-Arg-NH2; biotinyl-20 PHI (biotinyl-PHI-27), porcine; [Glp16] VIP 16-28, porcine; PHI (PHI-27), porcine; PHI (PHI-27), rat; PHM-27 (PHI), human; prepro VIP 81-122, human; preproVIP / PHM 111-122; prepro VIP / PHM 156-170; biotinyl-PHM-27 (bio-tinyl-PHI), human; intestinal vasoactive contractor (endothelin beta); vasoactive intestinal octacosa peptide, chicken; vasoactive intestinal peptide, capsule; biotinyl-VIP, human, porcine, rat; vasoactive intestinal peptide 1-12, human, porcine, rat; vasoactive intestinal peptide 10-28, human, porcine, rat; vasoactive intestinal peptide 11-28, human, porcine, rat, sheep; vasoactive intestinal peptide (cod, Gadus morhua); basic intestinal peptide 6-28; vasoactive intestinal peptide antagonist; vasoactive intestinal peptide antagonist (GHRF 1-29 [Ac-Tyr1, D-Phe2] -amide); vasoactive intestinal peptide receptor antagonist (4-CI-D-Phe6, Leu17] - VIP); and vasoactive intestinal peptide receptor binding inhibitor, L-8-.

Vasopressin peptides (ADH) including, but not limited to, vasopressin; [Asu 16, Arg 8] vasopressin; vasotocin; [Asu161Arg8] - vasotocin; [Lys8] -vasopressin; pressinoic acid; Desamino vasopressin [Arg] deglucinamide; [Arg] vasopressin (AVP); Vasopressin [Arg8] deglycinamide; biotinyl- [Arg8] vasopressin (biotinyl-AVP); [D-Arg8] - vasopressin; deamino [Arg8] vasopressin; deamino [D-Arg8] vasopressin (DDAVP); [deamino [D-3- (3'-pyridyl-Ala)] - [Arg8] vasopressin; [1- (beta-Mercapto-beta, beta-cyclopentamethylene propionic acid), 2- (0-methyl) tyrosine] - [Arg8] -vasopressin; vasopressin metabolite neuropeptide [pGIu4, Cys6]; vasopressin metabolite neuropeptide [pGIu4, Cys6]; [Lys8] - deamino vasopressin deglycinamide; [Lys8] -vasopressin; [Mpr1, Val4, DArg8] -vasopressin; [Phe2, He3, Orn8] -vasopressin ([Phe21 Orn8] -vasotocin); [Arg8] -vasotocin; and [d (CH 2) 5, Tyr (Me) 2, Orn 8] -vasotocin.

Virus-related peptides including, but not limited to, human fluorogenic CMV protease substrate; HCV 59-68 core protein; HCV NS4A 18-40 protein (strain JT); HCV NS4A 21-34 protein (strain JT); heptitis virus receptor binding fragment; hepatitis B virus pre-region 120-145; [Alal27] -pr-S hepatitus B virus region 120 - 131; herpes virus 2 inhibitor; HIV envelope protein fragment 254-274; HIV gag fragment 129-135; HIV substrate; P18 peptide; T peptide; [3,5 diiodo-Tyr7] T peptide; R15K HIV-I inhibitor peptide; T20; T21; V3 18-110 decapeptide P; and viral replication inhibitor peptide.

Wnt family proteins, and fragments thereof.

While certain analogs, fragments, and / or analogous fragments have been described from the various above polypeptides, it is to be understood that other analogs, fragments, and / or analogous fragments that retain all or some of the activity of the particular polypeptide, or otherwise act. as an antagonist thus impeding their action may also be useful in the embodiments of the present invention. Analogs can be obtained by various means, as will be appreciated by those skilled in the art. For example, certain amino acids may be substituted for other amino acids in a polypeptide without appreciable loss of interactive binding capacity with structures such as, for example, antibody antigen binding regions or binding sites on substrate molecules. As the interactive capability and nature of a polypeptide drug defines its biological functional activity, certain amino acid sequence substitutions may be made in the amino acid sequence and yet a polypeptide with similar properties remains, or otherwise confers this. analogous antagonistic activity with which it interferes or blocks the action of the natural product. In addition, small molecules, whether peptidomimetic or not, natural or synthetic, may be able to replace the proteins and peptides mentioned above and have similar activity by binding to their receptors. In contrast, such small molecules may block or interfere with the activity of the proteins and peptides cited above by various mechanisms, including, but not limited to, preventing their interaction with their cognate receptors. Additionally, many of the proteins cited above act as signaling pathway primers. One embodiment of this invention is the use of chemical molecules (peptides, peptidomimetics, or any other natural or synthetic molecule of any chemical nature) as activators or inhibitors of these signaling pathways. Examples of this strategy are the use of gamma secretase inhibitors to inhibit the Notch1 signaling pathway or inhibitors of the interaction between beta-catenin and Tcf transcription factors to inhibit the Wnt beta-catenin pathway. so involved in colorectal cancer. g) Oligonucleotide Agents

Active agents may also be in the form of oligonucleotides, including oligoribonucleotides, oligodeoxyribonucleotides and derivatives thereof useful for prophylactic, palliative or therapeutic purposes, including gene therapy and cancer treatment, such as colon cancer. An oligonucleotide is a polymer of a repeating unit generally known as a nucleotide. An unchanged (naturally occurring) nucleotide has three components: (1) a heterocyclic base containing nitrogen bound by one of its nitrogen atoms to (2) a 5-pentofuranosyl sugar and (3) a stereoisate phosphate in one of 5 'or 3' carbon atoms of sugar. When incorporated into an oligonucleotide strand, the phosphate of a first nucleotide is also esterified into an adjacent sugar of a second, adjacent nucleotide via a 3'-5 'phosphate bond. Nucleotides are nucleosides that also include a phosphate group covalently attached to the sugar moiety of the nucleoside. In oligonucleotide formation, phosphate groups covalently link adjacent nucleosides together to form a linear polymeric compound. The respective ends of this linear polymeric structure may also be joined to form a circular structure, but, within the context of the invention, open linear structures are generally preferred.

Oligonucleotides may include sufficient identity and number nucleotide sequences to perform specific hybridization to a particular nucleic acid. Such oligonucleotides that specifically hybridize to a portion of the sense filament of a gene are commonly described as "antisense". In the context of the invention, "hybridization" means hydrogen bonding, which may be Watson-Crick, Hoogsteen or reverse Hoogsteen hydrogen bonding, between complementary nucleotides. For example, adenine and thymine are complementary nucleobases that pair through the formation of hydrogen bonds. "Complementary" as used herein refers to the ability for precise pairing between the two nucleotides. For example, if a nucleotide at a certain position in an oligonucleotide is capable of hydrogen bonding with a nucleotide at the same position as a DNA or RNA molecule, then the oligonucleotide and DNA or RNA are considered complementary to each other at that position. . The oligonucleotide and DNA or RNA are complementary to each other when a sufficient number of corresponding positions in each molecule are occupied by nucleotides that can bond hydrogen to each other.

The term "oligonucleotide" refers to an oligomer or polymer of ribonucleic acid (RNA) or deoxyribonucleic acid (DNA) or mimetics thereof. This term includes oligonucleotides composed of naturally occurring nucleobase bases, sugars and covalent interchain sugars (main chain) as well as oligonucleotides having similarly functioning non-naturally occurring moieties. They can be single or bifilaments. In general, oligonucleotides formulated in the compositions of the invention may be from about 8 to about 100 nucleotides in length, more preferably from about 10 to about so nucleotides in length, and more preferably from about 10 to about 25 nucleotides. lengths.

Oligonucleotides that are formulated in the inventive compositions include antisense compounds and other bioactive oligonucleotides. A discussion of antisense oligonucleotides and some desirable modifications can be found in De Sameeker et al. (Acc. Chem. Res., 1995,28,366).

As used herein, antisense compounds include antisense oligonucleotides, antisense peptide nucleic acids (PNAs) 1 small interfering RNA, short pin RNA, ribozymes, and outer guide sequences (EGSs). In antisense messenger RNA (mRNA) modulation, hybridization of an antisense compound to its niRNA target interferes with the normal role of mRNA and causes a modulation of its function in cells. The mRNA functions to be interfered with include all vital functions such as RNA translocation to the protein translation site, current RNA protein translation, RNA splicing to yield one or more mRNA species, spinning or degradation of the mRNA. mRNA and possibly even independent catalytic activity that can be blocked by RNA. The general effect of such interference with mRNA function is modulation of a protein expression, where "modulation" means either an increase (stimulation) or a decrease (inhibition) in protein expression. In the context of the present invention, inhibition is the preferred form of gene expression modulation.

Antisense compounds can exert their effect by a variety of means. Such means are the antisense mediated direction of an endogenous nuclease, such as RNase H in eukaryotes or RNase P in prokaryotes, to the target nucleic acid (Chiang et al., J. Biol. Chem., 1991, 266, 18162. (Forsteretal., Science, 1990, 249, 783).

The sequences that recruit RNase P are known as External Guide Sequences, hence the abbreviation "EGS" (Guerier-Takada et al., Proc. Natl. Acad. Sci. USA, 1997, 94, 8468). Other means involve covalently linking a synthetic moiety having nuclear activity to an oligonucleotide having an antisense sequence, rather than recruiting an endogenous nuclease. Synthetic halves having nuclease activity include, but are not limited to, enzymatic RNAs, lanthanide ion complexes, and others (Haseloff et al., Nature, 1988, 334, 585; Bakeretal., J. Am. Chem. Soc. 1997, 119, 8749).

As used herein, the term "antisense compound" also includes ribozymes, synthetic RNA molecules, and derivatives thereof that catalyze highly specific endoribonuclease reactions (see generally US Pat. No. 5,543,508 to Haseloffet al. and US Pat. No. 5,545,729 to Goodchild et al.).

In addition, the term "antisense compound" includes RNA (or DNAs encoding such RNAs) leading to modulation of gene expression by the RNA interference mechanism. Such molecules include, but are not limited to, short interfering RNAs, consisting of bifiling RNAs of less than 50 base pairs, typically 21 or 29 nucleotides in length with the addition at either end of other chemical molecules. (including deoxyribonucleotides, natural or modified), as well as short-pin RNA (or DNA molecules including plasmids and viruses of any nature that lead to their production, in vitro or in vivo) that acts through interfering RNA. This also includes any single or double stranded DNA or RNA molecule that leads into cells for interfering RNA.

The antisense compounds formulated in the compositions of the invention (1) may be from about 8 to about 100 nucleotides in length, more preferably from about 10 to about 30 nucleotides in length, (2) single or bifilamentary, (3) are labeled for a nucleic acid sequence required for expression of a gene from a mammal, including a human, and (4), when contacted with cells expressing the target gene, modulate its expression. . Due to the biological activity of the gene product encoded by the target gene, modulation of its expression has the desirable result of providing specific prophylactic, palatable and / or therapeutic effects.

It is understood in the art that the nucleobase sequence of an oligonucleotide or other antisense compound need not be 100% complementary to its target nucleic acid sequence to be specifically hydrolysable. An antisense compound is specifically hydrolyzable to its target nucleic acid when there is a sufficient degree of complementarity to prevent nonspecific binding of the oligonucleotide to unlabeled sequences under conditions where specific binding is desired, that is, under physiological conditions in the environment. in vivo tests or therapeutic treatment, or in in vitro tests under test conditions.

Other bioactive oligonucleotides include aptamers and molecular callers. As used herein, the term is meant to refer to any oligonucleotide (including a peptide-nucleic acid or PNA) that (1) provides a prophylactic, palliative or therapeutic effect to an animal in need thereof and (2) acts by a non-antisense mechanism, that is, by some different means of hybridization to a nucleic acid. The name aptamer was created by EHington et al. (Nature, 1990,

346, 818) to refer to nucleic acid molecules that have adjusted and therefore bind with significant specificity to non-nucleic acid binders such as peptides, proteins and small molecules such as drugs and dyes. Because of these specific ligand binding properties, nucleic acids and oligonucleotides that can be classified as aptamers can be easily purified or isolated by affinity chromatography using immobilized ligand-bearing columns. Aptamers may be nucleic acids that are relatively short to those that are as large as a few hundred nucleotides. For example, RNA nucleotides that are 155 nucleotides in length and that bind dyes such as Cibacron Blue and Blue Reactive 4 with good selectivity have been reported (Ellington et al., Nature, 1990, 346, 818). Although RNA molecules were first referred to as aptamers, the term as used in the present invention refers to any nucleic acid or oligonucleotide that exhibits specific binding to small molecule ligands including, but not limited to, DNA, RNA, derivatives. DNA and conjugates, RNA derivatives and conjugates, modified oligonucleotides, chimeric oligonucleotides, and gapmers (see, for example, US Pat. No. 5,523,3B9, to Ecker et al., issued June 4, 1996 and incorporated here by reference).

Molecular decoys are short bifilamentary nucleic acids (including single-stranded nucleic acids designed to "fold back" on themselves) that mimic a site in a nucleic acid to which a factor, such as a protein, binds. Such decoys are expected to competitively inhibit the factor; that is, because the factor molecules are bound to an excess of the decoy, the cell site-bound factor concentration corresponding to the decoy decreases, with resulting therapeutic, palliative or prophylactic effects. Methods of identifying and constructing decoy molecules are described, for example, in U.S. Pat. No. 5,716,780 to Edwards et al.

Another type of bioactive oligonucleotide is a RNA-DNA hybrid molecule that can direct gene conversion of an endogenous nucleic acid (Cole-Strauss et al., Science, 1996, 273, 1386).

Preferred modified oligonucleotide backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorothithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3'-alkylene phosphonates and chiral phosphonates, phosphonates, phosphoramidates including phosphoramidates -amino and aminoalkylphosphoramides, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates having normal 3'-5 'bonds, analogues 2'-5' linked thereof, and those having inverted polarity in which adjacent pairs of nucleoside units 3 '-5' to 5-3 'or 2'-5' are linked to 5'-2 '. Various salts, mixed salts and free acid forms are also included. Any of the foregoing bioactive oligonucleotides may be formulated in the drug delivery system of the invention and used for prophylactic or therapeutic purposes. Oligonucleotides may be stabilized by complexing, for example, with cationic lipids such as Lipoplexe or cationic polymers such as Poliplexe. h) Diagnostic Agents

Medical imaging is noninvasive or non-surgical visualization of internal organs or processes. Representative diagnostic methods include X-rays, magnetic resonance imaging (MRI), radionuclides or nuclear medicine, and ultrasound. Radionuclides are nuclei that deteriorate upon dissolution.

excess energy (parent) to become stable (child) by emission of energy in the form of particulate or electromagnetic radiation. Fluoroscopy is a fluorescent screen that detects gamma or X-rays that are transmitted by a TV camera to provide real-time images of> 20 moving organs using contrast agents such as PCTA. CAT - Axial computed tomography - takes advantage of small differences in radiographic tissue density to create an image. The colon is often imaged using a lower GI series of a barium enema to conduct a radiographic study of the large intestine colon and rectum. Technetium is a common radiolabel. Other radioactive compounds

Labels include iodine radiolabels such as iobenanol sulfate 131I1 sodium iodine123, sodium iodine, and indium labels such as 111In radiolabels, indium chloride, and indium satumomabpendetide. Imaging contrast agents include iron-containing contrast agents such as ferumoxides and dentritic gadolinium.

The present invention may be used to release to colon agents that allow or facilitate visualization of structures, lesions, cells carrying cell surface or intracellular molecules defined by any imaging technique including, but not limited to, radiography, radiotomography, imaging magnetic resonance imaging (MRI) 1 ultrasound, positron emission tomography (PET scan), or any other form of imaging technique using radiomagnetic waves of any wavelength. For example, small molecules or antibodies that recognize cell surface structures of colon cancer cells may be radionuclide labeled such as "Technetium" and used to detect tumor cells and metastasis of various sizes including micrometastases.

II. METHODS TO PREPARE PECTIN ACCOUNTS

Pectin beads can be prepared using methods known to those skilled in the art, including mixing the active agent (s) in a pectin solution, and gelling the anionic halves of ketine with a divalent cation. such as divalent zinc, for example in the form of a zinc acetate solution.

Gelling is typically done by stirring a solution, suspension or dispersion of the active agent in one embodiment, β-lactamase L1, and pectin, adjusting the pH of the solution if necessary, and adding this solution dropwise to a solution of. stirring zinc acetate. In some embodiments where the active agent (s) are not adversely affected by other metal ions, divalent or trivalent metal ions other than zinc may be used.

Suitable technologies for adding the pectin drop solution to the zinc acetate solution are known to those skilled in the art; and include Nisco Engineering AG's multi-tap system and other relevant technologies for producing drops of a solution of petin.

Pectin drops undergo a gelling process, ideally for a predetermined time to obtain the best encapsulation yield and subsequent release efficiency.

The concentration of the pectin solution is advantageously about 4 to about 10% (w / v), preferably about 4 to about 7%, the metal cation such as zinc acetate solution. It is advantageously from about 2 to about 20% (w / v), preferably from about 5 to about 15%. More preferably, the pectin solution is about 5% (w / v), the zinc acetate solution is about 12% (w / v).

The pectin beads are advantageously stirred in the metal cation, such as zinc acetate solution, at a pH of about 6 at room temperature, under slow stirring, for at least about 12 minutes until around 20 hours, preferably around 20 minutes to around 2 hours.

The beads can then be collected and rinsed in distilled water, ideally until the conductivity of the rinse solution reaches a plateau. Rinsing is preferably done at least twice or under a continuous process to minimize the amount of residual zinc acetate if restored in the rinsing solution.

The rinsed beads may then be collected and subjected to a drying process using methods known to those skilled in the art, including heated or fluidized bed incubator technologies.

The beads are typically dried at a temperature of between

between about 20 and about 40 ° C for about 30 min at about 24 hours, preferably at around 35 ° C overnight. Drying is preferably performed until the weight of the beads reaches a plateau.

The particle diameter can be finely tuned using needles of appropriate inner diameter to form the pectin droplets added to the zinc acetate solution. The beads are preferably between about 600 and 1500 pm in diameter.

When the active agent is β-lactamase L1, encapsulation yields are typically between 50 and 100%, measured in terms of enzymatic activity.

III. FORMATION OF DRUG RELEASE SYSTEMS INCLUDING PECTIN ACCOUNTS Pectin beads can be collected, and combined with appropriate excipients and formulated in a variety of oral drug release systems. For example, the beads may be combined with a solid, tableted excipient or enclosed in a capsule.

Pectin beads may also be combined with liquid / gels that do not degrade pectin beads, and the mixture / dispersion may be incorporated into a capsule, such as a gel capsule.

The tablets or capsules may be coated, if desired, with a suitable enteric coating to assist in crossing the stomach without degradation. The pH in the stomach is on the order of 1 to 3, but it increases in the small intestine and colon to reach values close to 7 (Hovgaard L. et al. (1996). Current Applications of Polysaccharides in Colon Taringing, Critical Reviews in Therapeutic Drug Carrier Systems, 13, 185). Drug release systems, in the form of tablets, gelatin capsules, spheroids and others, can reach the colon without being exposed to these pH variations, coating them with a pH-dependent polymer, acidic but pH-soluble insoluble. neutral or alkaline pH (Kinget et al. op. cit). The most commonly used polymers for this purpose are methacrylic acid derivatives, Eudragit® LeS (Ashford M. et al. (1993), An in vivo investigation of the suitability of pH-dependent polymers for colonic targeting, International Journal of Pharmaceutics, 95 , 193 and 95, 241; and David A. et al (1997) Acrylic Polymers for Colon-Drug Delivery, STP Pharmaceutical Sciences, 7, 546) and more recently Eudragit® FS.

Drug delivery systems are administered in an effective amount adequate to provide the appropriate degree of treatment or prevention of the disorders for which the compounds are administered. The efficient amounts of these compounds are typically below the threshold concentration required to elicit any appreciable side effect. The compounds may be administered in a therapeutic window in which some disorders are treated and certain side effects are avoided. Ideally, the effective dose of the compounds described herein is sufficient to provide the desired effects on the colon but is insufficient (that is, not at a high enough level) to provide undesirable side effects elsewhere in the body.

More preferably, effective doses are at very low concentrations, where maximum effects are observed to occur, with minimal side effects, and these are optimized by marked colonic release of the active agents. Prior effective doses typically represent that amount administered as a single dose, or as one or more doses administered within a 24 hour period. IV. TREATMENT METHODS USING THE PHARMACEUTICAL RELEASE SYSTEMS DESCRIBED HERE

The drug delivery systems described herein may be used to treat those types of conditions and disorders for which colonic release is appropriate. In one embodiment, the disorders are those that result from exposure of the colon to antibiotics such as diarrhea, modification of the commensal flora, and the development of bacterial resistance to antibiotics. In this embodiment, drug delivery systems contain antibiotic inactivating agents, and the active ingredients may be administered in a therapeutically effective dosage to a patient who has been, is, or will be treated with one or more antibiotics.

Where the metallo-dependent enzyme is an enzyme other than one that inactivates antibiotics, such an enzyme may be administered to treat the specific disorders treated by such enzymes.

In another embodiment, drug delivery systems are administered to a patient suffering from colon cancer. In this embodiment, drug delivery systems include one or more anti-tumor agents, and the systems are administered at a therapeutically effective dosage to a patient suffering from colon cancer. Alternatively, cancer may be present elsewhere in the body, and drug delivery systems may be used to bypass the stomach and its concomitant degradation of certain antitumour agents to avoid the need for intramuscular or intravenous administration. of these agents.

In another embodiment, drug delivery systems are administered to a patient suffering from a colonic disorder such as Chrohn's disease ulcerative colitis, irritable bowel syndrome, diarrhea, or constipation. In this embodiment, drug delivery systems include agents that treat or prevent such disorders, and the systems may be administered in a therapeutically effective dosage to a patient experiencing such disorder.

In yet another embodiment, drug delivery systems are used to deliver peptide or protein-based active agents such as insulin, antibodies, and others, or oligonucleotide-based therapies such as antisense therapy or interfering RNA. , so that agents pass through the stomach undigested. In this embodiment, drug delivery systems include these protein / peptide / oligonucleotide-based agents, and the systems may be administered at a therapeutically effective dosage to a patient in need of treatment with these agents without the need for additions. - administer these agents by subcutaneous or intravenous injection.

In another embodiment, drug delivery systems are used to deliver diagnostic agents to the colon. In this fashion, drug delivery systems include diagnostic agents, such as imaging contrast agents, and the systems are administered at a diagnostically effective dosage to a patient who will undergo a diagnostic assay for the diagnosis of a colonic disorder.

The present invention will also be understood with reference to the following non-limiting examples.

EXAMPLE 1: DEVELOPMENT OF A SENSITIVE, QUANTITATIVE AND SPECIFIC TEST FOR β-LACTAMASE L1

Nitrocefin hydrolysis is a well known technique used to quantify penicillinase activity. However, the usual format is in single tubes and is not adapted for analysis of a large number of samples. This example describes the development and suitability for qualification of this 96-well microplate format assay.

A nitrocefin feedstock solution was obtained by dissolving dry nitrocefin powder at a concentration of 10 mM in dimethyl sulfoxide (DMSO). The feedstock solution was stored at -20 ° C and diluted 100 times immediately before use in 50 mM sodium phosphate buffer (Hepes buffer) pH 7.0 containing 0.1 mg / ml bovine serum albumin ( BSA). Buffer selection is described in Table I.

μΙ containing the solution to be analyzed were added to 180 μΙ of diluted nitrocefin. Nitrocefin hydrolysis kinetics were followed at 37 ° C with absorbance measured at 492 nm every 30 seconds using a Multiskan Ascent plate reader (Thermo Labsystems).

The slope (difference in absorbance / second) was calculated using Excel Adds In Cellula (Prisma Technologies, Cambridge UK). Lactamase L1 (Eurogentec, Belgium, approx. 10 mg / mL were determined by pBCA assay) was diluted 500x, 1000x, 2000x and 4000x in each solubilization buffer and the reaction was started by adding 20 μΙ of enzyme-containing solution to 180 μΙ buffers containing nitrocefin at 100 μΜ.

Β-lactamase L1 activity was tested in 10 mM Hepes, 145 mM NaCl buffer pH 7.4. The interference of EDTA with metallo-dependent enzyme activity and the need for a vehicle protein (Bovine Serum Albumin, abbreviated as BSA) was tested. As illustrated in Table 2, EDTA (which can be used to solubilize beads in vitro to test their contents) should be avoided. The inclusion of BSA or other carrier proteins is beneficial.

TABLE 2: SELECTION OF BUFFER COMPOSITION FOR QUANTIFIED B-LACTAMASE L1 ACTIVITY

Buffer_Declive Yield

10mM Hepes, 145mM NaCl pH 7.4 0.142 100%

mM Hepes, 145 mM NaCl, 1% EDTA pH 7.4 0.026 18.8% mM Hepes, 145 mM NaCl, 0.1 mg / ml BSA pH 7.4 0.167 118.2% (mM Hepes, 145 mM NaCl, 0.1 mg / ml BSA, 1% 0.084 59.0% EDTA pH 7.4

As shown in Table 1, EDTA interferes with the enzymatic activity assay, and BSA enhances the restoration of enzymatic activity.

Example 2: β-LACTAMASE L1 INSTABILITY IN ORIGINAL PECTIN MIXTURE AND METAL COUNTER ION EFFECT

0.3 ml of β-lactamase L1 (Eurogentec, Belgium, approx. 10mg / ml as determined by the pBCA assay) was mixed with 10g of a 6% solution of pectin (low methoxylated amidated pectin (Unipectine) 1 Tex - turant Systems, cat # OG175C) made in water; The pH of the pectin solution was not adjusted.

The pectin / β-lactamase L1 mixture was added dropwise over a 2 minute period using a peristaltic pump and a 0.8 mm internal diameter needle to a beaker containing 40 ml of calcium chloride (6%) while stirring. (200 rpm) at room temperature.

To allow equilibrium between the free and bound calcium ions, the beads were restored by filtration after further incubation and washed 3 times in 200 ml of purified water to eliminate excess free calcium. At this stage, the beads are referred to as "gelled beads".

The beads were dried 2 hours at 37 ° C in an oven, yielding dried beads. 2x5 droplets and 2 χ 15 droplets were sampled at the needle outlet to measure the initial activity of β-lactamase L1. Protein free beads were also prepared as negative controls. The enzymatic activity of β-lactamase L1 (nitrocefin hydrolysis) was quantified with and without Zn ions (0.1 mM ZnCl2) as described in example 1.

As shown in Table 3, no enzymatic activity was found in the β-lactamase L1 / pectin mixture while significant activity was restored to the beads tested in the buffer containing Zn + 2. TABLE 3: Inactivation of β-LACTAMASE L1 IN SOLUTION OF UNPUNTED PECTIAN IN pH

Decline / min_Sem ZnAc With ZnAc_

Prior to mixing with pectin 0.105 0.103 (100.0%) (100.0%) β-Lactamase / pectin mixture 0.000 0.000

(0.0%) (0.0%) Gelled milli particles 0.0078 0.042

_ (7.4%) _ (40.7%) _

EXAMPLE 3: OPTIMIZATION OF METAL ION USED TO GEL PECTIN, AND THE EFFECT OF PH OF PECTIN SOLUTION.

To determine the effects of pectin and zinc ion solution parameters, an experiment comparing four formulations was performed. The project was built according to factorial design, Design Ex- pert 6.0.10, Stat-Ease, Minneapolis. Two parameters were tested:

(a) pH of pectin solution: 4.0 and 7.0

(b) the metallic cation in the gelling bath: Ca2 + (CaCl2) or Zn2 + (zinc acetate abbreviated ZnAc)

Beads were prepared as described in Example 2. However, the concentration of pectin solution was decreased from 6% to 4% due to the decrease in pectin solubility with increased pH.

Encapsulation yield was measured by assaying the enzymatic activity of β-Lactamase L1 as described in Example 1.

The beads were solubilized in 20 ml of 10 mM Hepes, 145 mM NaCl, 0.1 mg / ml BSA at pH 7.4 in the presence or absence of 1% pectinase (Aspergillus Aculeatus pectinases, Pectinex SP- L Ultra (SIGMA, France) overnight at 4 ° C.

The positive control was prepared by diluting the same amount of β-lactamase L1 as it would contain in 5 beads in 20 ml of 10 mM Hepes, 145 mM NaCl, 0.1 mg / ml BSA pH 7.4. As shown in Table 4, β-lactamase L1 was inactivated regardless of the cation used for pectin gelation when the pectin solution was at pH 4.0 (4.3% of residual calcium activity and 3.8% zinc). ), while almost total activity was retained after buffering the pectin solution to pH 7.0 (86.7% calcium and 64.0% zinc).

TABLE 4: Effect of cation used for gelation and pH of pectin on stability (restoration of 3-lactamase activity)

Sample CaCl2, pH4 CaCl2, pH7 ZnAc5 pH4 ZnAc, pH7 Prior to mixing 0.102 0.090 0.108 0.090 (100%) (100%) (100%) (100%) Gelled Beads 0.004 0.072 0.004 0.072 Ov CO (80.0%) ( 3.8%) (80.0%) Dried accounts 0.003 0.078 0.037 0.058 (31.7%) (86.7%) (35.0%) (64.0%)

EXAMPLE 4: DETERMINING CRITICAL PARAMETERS FOR FORMULATING β-LACTAMASE L1 FOR COLON-SPECIFIC RELEASE

AND OPTIMIZATION OF THESE PARAMETERS Five parameters were tested:

(a) Concentration of pectin solution (Low Methylated Amide Pectin (Unipectine), Texturant Systems, cat # OG175C): 4% and 5% (w / v)

(b) Gelation cation: Ca2 + or Zn2 +

(c) Secondary coating of the gelatin beads with solution.

Polyethyleneimine (PEI) (PEI, High Molecular Weight, Water Free (SIGMA-ALDRICH, France))

(d) pH of PEI solution: 7 and 11 (original solution not adjusted

at pH).

(e) Solubilization of beads to test enzymatic activity

encapsulated with and without 1% pectinase.

Table 5 summarizes the experimental design TABLE 5: EXPERIMENTAL DESIGN FOR THE OPTIMIZATION OF CRITICAL PARAMETERS INVOLVED IN FORMULATION OF β-LACTAMASE L1 Operation A: Pectin B: Ion: D: pH of E: Peetina- (%) PEI if

PEI

1 5 Zn2 + Yes 11 Yes 2 4 Zn2 + Yes 7 Yes 3 5 Ca2 + No Yes 4 4 Ca2 + Yes 7 No 5 Ca2 + Yes 7 Yes 6 4 Ca2 + No Yes 7 4 Zn2 + Yes 11 No 8 5 Ca2 + Yes 11 No 9 4 Ca2 + Yes 11 Yes 4 Zn2 + No No 12 4 Zn2 + No Yes 13 5 Zn2 + Yes 7 No 14 5 Ca2 + No No 4 Ca2 + No No 16 5 Zn2 + No No

These 16 experiments were performed in duplicate (32 results). Operation 13 replicated (34 results).

The pH of 4% and 5% of pectin solutions was adjusted to 7.0. However, it was determined that the pH of the 5% pectin solution was unstable and decreased to pH 5.4 at the end of the experiments. A 5% pectin solution was therefore also adjusted to pH 8.5 for comparison.

Finally, the 48 results were analyzed using Project Factorial

rial.

The beads were prepared as described in Example 2 except that the gelation time in the cation bath was reduced from 20 min to min to allow intelligent regulation of the experiments.

Samples (5 beads) were solubilized overnight at 4 ° C in 20 ml of 10 mM Hepes, 145 mM NaCl, 0.1 mg / ml BSA pH 7.4 with and without 1% pectinase before measuring. enzymatic activity (nitrocefin hydrolysis as described in example 1).

Table 6 summarizes the experimental results obtained. TABLE 6: TOTAL RESULTS OF THE EXPERIMENTAL DESIGN TO OPTIMIZE CRITICAL PARAMETERS INVOLVED IN THE FORMULATION OF 3-LACTAMASE L1 PERIOD% PH ION PEI PH Pectinase PETININE YIELD PEI 1 5 5.4 Zn2 + yes 11 yes 1.201 17 5 4 Zn2 + yes 11 yes 1.13 3b 5 5.4 Zn2 + yes 11 yes 1.39 11 5 5.4 Zn2 + no yes 1.272 27 5 5.4 Zn2 + no yes 1.36 2b 5 5.4 Zn2 + no yes 1.044 1b 5 5.4 Zn2 + yes 7 yes 1.045 13 5 5.4 Zn2 + yes 7 no 0.687 29 5 5.4 Zn2 + yes 7 no 0.72 33 5 5.4 Zn2 + yes 7 no 0.661 34 5 5.4 Zn2 + yes 7 no 0.691 16 5 5.4 Zn2 + no no 0.762 32 5 5.4 Zn2 + no no 0.788 45 5 8.5 Zn2 + no yes 0.951 38 5 8.5 Zn2 + no yes 0.818 41 5 8.5 Zn2 + no no 0.245 48 5 8 5 Zn2 + no no 0.363 46 5 8.5 Zn2 + yes 7 no 0.815 39 5 8.5 Zn2 + yes 7 no 0.826 2 4 7 Zn2 + yes 7 yes 1.01 18 4 7 Zn2 + yes 7 yes 1.162 12 4 1 Zn2 + no yes 1.165 28 4 1 Zn2 + no yes 1.118 7 4 7 Zn2 + yes 11 no 0.727 23 4 7 Zn2 + yes 11 no 0.679 4 7 Zn2 + no no 0.674 26 4 7 Zn2 + no no 0.659 3 5 5.4 Ca2 + yes 7 yes 0.094 5 5.4 Ca2 + yes 7 yes 0.031 Operation% PH ion PEI PH pectinase specific yield PEI 19 5 5.4 Ca2 + yes 7 yes 0.108 21 5 5.4 Ca2 + yes 7 yes 0.039 8 5 5.4 Ca2 + yes 11 no 0.047 24 5 5.4 Ca2 + yes 11 no 0.066 14 5 5.4 Ca2 + no no 0.488 5 5.4 Ca2 + no no 0.512 5 8, 5 Ca2 + yes 7 yes 0.35 36 5 8.5 Ca2 + yes 7 yes 0.379 42 5 8.5 Ca2 + yes 7 yes 0.363 43 5 8.5 Ca2 + yes 7 yes 0.394 4b 5 8.5 Ca2 + yes 7 yes 0.53 7b 5 8.5 Ca2 + yes 7 no 0.704 37 5 8.5 Ca2 + yes 11 no 0.029 44 5 8.5 Ca2 + yes 11 no 0.029 9b 5 8.5 Ca2 + yes 11 no 0.737 40 5 8.5 Ca2 + no 0.322 47 5 8.5 Ca2 + no 0.656 6b 5 8.5 Ca2 + yes 11 yes 0.517 5b 5 8.5 Ca2 + no yes 0.656 8b 5 8.5 Ca2 + no no 0.967

Simple monovariate statistical analysis (decreasing yield

encapsulation classification) highlighted that an optimal β-lactamase L1 formulation was obtained using the following parameters:

(a) A pectin concentration of 5% (maximum solubility

at pH 5.4)

(b) A neutralized pectin solution at a pH of at least 5.4

(c) A zinc ion should be used.

(d) A secondary coating may also be evaluated with another type of polymers.

(e) A pectinase should be used to quantify formulated β-lactamase L1

EXAMPLE 5: IMPROVING ACCOUNTING STABILITY UNDERSTANDING 3-LAETAMASE L1 IN SIMULATED INTESTINAL MEANS (YES) FOR INCREASED ZINC ION CONCENTRATION AND DRYING DURATION

Beads containing β-lactamase L1 were prepared as described in example 4. Increasing zinc acetate concentrations (6, 8, 10 and 12%) were tested. Another coating with or without PEI was compared.

Drying of the beads was also increased from 2 hours to overnight. Washing efficiency to remove excess metal ion used for gelation was also monitored by measuring the conductivity of the water rinse solution.

As illustrated in Figure 1, efficient washing was obtained after washing the beads in three water washes.

As shown in Table 7, the higher concentration of zinc acetate increased SIM (Simulated Intestinal Medium, US Pharmacopeia 26) stability of β-lactamase L1-containing beads while secondary PEI coating decreased their stability.

TABLE 7: EFFECT OF ZINC ACETATE CONCENTRATION AND SECOND PEI COATING ON THE STABILITY OF ACCOUNTS CONTAINING Lβ-LACTAMASE L1 IN YES_

YES Operation N0% Zn PEI 1h 2h 3h 4h 5h 8 10% N + + + + + 3 12% S + + + + + 4 12% N + + + + + 8 8 N + + + + + 6% S - - - - - 1 8% S + + - - - 7 10% S + - - - - +: stable beads -: dissolved beads S: with PEI backing N: without PEI backing

EXAMPLE 6: EFFECT OF ZINC CONCENTRATION AND DRYING TIME ON STABILITY OF ACCOUNTS IN SIMILAR INTESTINAL MEANS (YES)

Beads containing β-lactamase L1 were previously prepared as described, and gelled with 6 or 12% zinc acetate solutions (see Example 5).

The effect of drying time was also tested by drying the beads for 2, 4 and 16 h at 35 ° C (preferred temperature at 37 ° C for industrialization purposes). Only beads gelled in the 12% zinc solution and dried for more than 4 h were stable in YES after 5 h incubation at 37 ° C. The stability of YES beads for 5h @ 37 ° C was measured, and the results are shown in Table 8. TABLE 8: BALANCE STABILITY IN SIMILAR INTESTINAL MEDIA FOR 5H AT 37 ° C. NUMBERS STILL REPRESENT NUMBER OF ACCOUNTS APPEARANCE INTACT IN SOLUTION._

Incubation at 2 h of drying 4 h of drying at 37 ° C (h) Gem night milli 1 5 0 0 particles 2 1 0 0 in 5% of 3 1 0 0 Zn 4 1 0 0 5 1 0 0 milli- 1 5 5 5 particles 2 5 5 5 in 12% 3 5 5 5 of Zn 4 4 5 5 5 3 4 5

After washing and additional incubation in simulated colonic medium (SCM): 10 mM Hepes, 145 mM NaCl (raw material solution). 1% pectinase, 0.1 mg / ml BSA were added just prior to use; pH was adjusted to pH 6.0 with 1 M NaOH, 63% of the initial activity of β-Iactamase (nitrocefin hydrolysis) was restored. EXAMPLE 7: EFFECT OF FREEZING TIME, RINSE PROCESS, AND DRYING TIME UNDER RESTORATION OF β-LACTAMASE L1 ACTIVITY

Different batches of beads were prepared using an Engeneering AG Nisco ATM system. The beads have undergone several times gelation, rinsing process and time and type of drying process and time.

It clearly appeared that the best encapsulation efficiency and enzyme activity is obtained when the gel time is less than hours and when rinsing is performed such as to eliminate residual Zinc acetate from the beads. The results are shown in Figure 2, <■ EXAMPLE 8: DEVELOPING A SENSITIVE, QUANTITATIVE AND SPECIFIC TEST FOR β-LACTAMASE L1

CENTA Hydrolysis is a well known technique used to qualify β-lactamase activity. However, the usual format is in single tubes and is not adapted for analysis of a large number of samples. This example describes the development and fit for qualification purposes of this 96-well microplate format assay.

A solution of CENTA raw material was obtained by solubilizing the dry CENTA powder to a concentration of 25 mM in water; It was stored in 25 μΙ aliquots at -20 ° C. The assay mixture was terminated by diluting 22 μΙ of CENTA raw material solution in the following assay buffer: 10 ml of 30 mM Hepes pH 7.5 buffer containing 50 pm ZnCl2, thus yielding a concentration of CENTA of 110 μΜ. For the assay, 20 μΙ containing the enzyme to be tested were added to 180 μΙ of the assay mixture, consequently using a final concentration of 100 μΜ of CENTA in the assay. CENTA hydrolysis kinetics were followed at 37 ° C with an absorbance measurement at 405 nm every 9 seconds using a Multiskan Ascent plate reader (Thermo Electron Corporation). The slope (difference in absorbance / second) was calculated using Ascent Software for Multiskan Ascent version 2.6.

β-lactamase L1 (Eurogentec, Belgium, approx. 10 mg / mL as determined by the pBCA assay) was diluted to 0.2, 0.5, 1.0 and 2.0 pg / mL in assay buffer and the reaction was initiated by the addition of 20 μΙ of solution containing the enzyme at 180 μΙ of the assay mixture. As shown in the Figure below, the assay was linear in 3 independent assays with respect to enzyme concentration in that range. Standard deviation was less than 10%. EXAMPLE 9. β-LACTAMASE L1 RELEASE FROM UN-COATED ACCOUNTS, AND EUDRAGIT COATED ACCOUNTS WITH OR WITHOUT HPMC PRE-COATING. A batch of β-lactamase L1-containing pectin beads was

manufactured under the following conditions: beads were formed by dropping through a 0.5 mm internal diameter needle a% pectin solution containing 300 mg / l purified recombinant β-lactamase L1 (Eurogentec, Belgium) in a 12% Zn1 acetate 2H20 bath. Beads were gelled for 90 min in the Zn acetate bath, collected, washed with water until the water conductivity reached a stable plateau, meaning that rinsing is optimal and finally dried at 35 ° C under vacuum. The dried beads obtained were 0.8 - 1.25 mm in diameter, weighed on average 0.6 mg and contained approximately 5 to 6 pg β-lactamase L1 per mg beads. They were left uncoated, or coated using a Glatt GPC 1.1 with TOP spray according to the following formulas shown in Table 9.

TABLE 9

Crude materials Quantity (g) Batch 83 Quantity (g) Batch 100 Quantity (g) Batch 82 Quantity (g) Batch 99 Quantity (g) Batch 81 Quantity (g) Batch 97 Eudragit L30D- 55 1600.0 149.5 300.0 31.9 Eudragit NE 30 D 700.0 74.4 Eudragit FS30D 800.0 85.0 GMS (glycerol monosearate) 24.0 2.2 15.0 1.6 12.0 1.3 Sodium hydroxide 28.8 2.7 30.4 1.9 1.5 Tween 80 48.0 2.2 18.0 1.6 14.4 1.3 (polysorbate) 33% aqueous solution Triethyl citrate 1107.2 94.5 4.50 67.2 10.0 25.2 Water 1600.0 149.5 565.7 1600.0 505.6 85.0 5% HPMC coating NO YES NO YES NO YES

Bead precoating was performed with HPMC using the same material as for Eudragit coating. 1 Electron Scanning Micrographs (SEMs) of coated beads

Eudragit shells are shown in Figure 4. A cross section shows the relative thickness of the Eudragit coating.

To assess β-lactamase L1 release, coated and uncoated beads were incubated under gentle mixing at 37 ° C in 50 mM Hepes pH 7.4 buffer containing 0.1 M NaCl and 100 PG / ml pectin - Aspergillus aculeatus (Sigma Aldrich) months. Medium was withdrawn at various times and assayed for β-lactamase activity using the nitrocepine assay described in Example 1. Release kinetics were measured using the coated beads and

uncoated, and the results are shown in Figure 5. EXAMPLE 10: EFFICIENCY OF L1 RELEASED TO ANTIBIOTIC HYDROLYSIS

To assess whether coated beads could actually hydrolyze antibiotics when they reached the colon, they were incubated successively for 1 h in simulated gastric medium (0.1 N HCI), 3 h at 37 ° C in simulated intestinal medium (50 mM Na / K phosphate buffer pH 6.8 containing 0.1 M NaCl) and finally for the indicated amounts of time in the simulated colonic medium (50 mM Hepes buffer pH 7.4, 0.1 M NaCl ) containing 100 µg PG / ml Aspergillus aculeatus pectinases (Sigma Aldrich) and 2 mg / ml amoxicillin. Medium was removed at various times and the amount of residual amoxicillin was measured by HPLC and UV absorption. The procedure was performed using a Bio-Diss Ill apparatus (Varian). Uncoated beads were only incubated in the simulated colonic medium with pectinases and amoxicillin.

The results are shown in Figure 6.

Example 11: Effect of β-lactamase L1 containing beads on the appearance of bacterial resistance in amoxicillin-treated guinea pig.

Piggy 6-7 weeks old or untreated or treated orally with 20 mg / kg amoxicillin per day for 7 days. Half of the treated animals received, along with the daily dose of antibiotics, a gelatin capsule filled with 320 mg pectin beads containing 5% HPMC pre-coated β-Iactamase L15 and coated with 40% Eudragit L30D- 55 (batch 100); the other half similarly received placebo-coated pectin accounts. Faeces were collected 3 days prior to treatment initiation, and each day for 7 days of treatment and analyzed for their total and amoxicillin-resistant enterobacterium content on MacConkey agar plates containing 0 or 100 Mg / ml amoxicillin. As shown in Figure 7, untreated animal feces contained a minimal proportion of amoxicillin-resistant bacteria (<5%), while this proportion rapidly increased in amoxicillin-treated animals, reaching a value between 50 and 80% after 7 days. In contrast, animals receiving β-lactamase-containing beads along with amoxicillin showed only a transient and limited increase in antibiotic-resistant bacteria. This experiment shows that the co-administration of the Eudragit-coated pectin beads containing β-lactamase L1 protected the guinea pig against the appearance of antibiotic resistant bacteria induced by treatment of animals with amoxicillin.

All patents and publications disclosed herein are incorporated by reference in their entirety. Modifications and variations of the present invention will be obvious to those skilled in the prior art detailed description of the invention.

Claims (18)

1. A drug delivery system for oral administration and colonic release of a prophylactic, therapeutic or diagnostic agent comprising a pectin bead containing a prophylactic, therapeutic or diagnostic agent wherein the pectin is cross-linked with a cation of metal and the bead is coated with a methacrylic acid-acrylic acid copolymer for enteric coatings or a polymer derived from esters of acrylic acid and methacrylic acid for enteric coatings, and wherein said agent is a protein, a nucleic acid , a virus, a bacterium, a fungus, a microorganism, a eukaryotic cell, and any combination thereof. The drug delivery system of claim 1, wherein said agent is selected from the group consisting of lipoproteins, glycoproteins, lipopeptides, glycopeptides, enzymes and oligonucleotides. 3. A drug delivery system for oral administration and colonic release of a prophylactic, therapeutic or diagnostic agent comprising a pectin bead containing a prophylactic, therapeutic or diagnostic agent wherein the pectin is crosslinked with zinc and the bead is coated with a methacrylic acid-alkyl acrylate copolymer for enteric coatings, or a polymer derived from esters of acrylic acid and methacrylic acid for enteric coatings. The drug delivery system of any of claims 1-3, wherein the agent is an agent that inactivates an antibiotic in the colon. The drug delivery system of any one of claims 1-3, wherein the agent is an anticancer agent, an anti-inflammatory agent, or a protein or peptide. The drug delivery system of any of claims 1-3, wherein the agent is an agent capable of treating a colon disorder, an agent that blocks or modulates the activity of receptors in the colon, or an agent. which inactivates other therapeutic agents that could modulate receptor activity in the colon. The drug delivery system of claim 4, wherein the antibiotic inactivating agent is a beta-lactamase, an erythromycin esterase, an enzyme inactivating a quinolone antibiotic or an inactivating enzyme. a glycopeptide antibiotic. The drug release system of claim 7, wherein the agent which inactivates an antibiotic is Stenotrophomonas maltophilia beta-lactamase L1. A drug delivery system according to any one of claims 1 to 8, wherein the methacrylic acid alkyl acrylate copolymer or the polymer derived from acrylic acid and methacrylic acid esters for enteric coatings is selected from the following: group consisting of alkyl methacrylic acid-acrylate copolymers or polymers derived from the esters of acrylic acid and methacrylic acid for enteric coatings with acidic or alkaline groups, or is selected from the group consisting of methacrylic acid-acrylate copolymers of alkyl or polymers derived from acrylic acid and methacrylic acid esters for alkaline or neutral enteric coatings. A drug delivery system for releasing a metallo-dependent enzyme, comprising pectin beads encapsulating the metallo-dependent enzyme, wherein the metal cation on which the enzyme depends is also used to crosslink the pectin, and wherein the ketone beads are coated with a methacrylic acid-alkyl acrylate copolymers for enteric coatings or a polymer derived from acrylic acid and methacrylic acid esters for enteric coatings. The drug delivery system of claim 10, wherein the metallo-dependent enzyme is beta-lactamase L1 from Steenrophomonas maltophilia. A drug delivery system according to claim 10 or 11, wherein the methacrylic acid-alkyl acrylate copolymer or polymers derived from acrylic acid and methacrylic acid esters for enteric coatings is selected from the group consisting of: methacrylic acid-alkyl acrylate copolymers with acidic or alkaline groups, or is selected from the group consisting of alkyl methacrylic acid-acrylate copolymers or polymers derived from acrylic acid and methacrylic acid esters for enteric coatings with alkaline or neutral groups. A drug delivery system according to any one of claims 1 to 4 and 7 to 12 for inactivating an antibiotic in a patient's colon. Drug release system according to any one of claims 1 to 6 and 9, for the treatment of disorder such as Crohn's disease or ulcerative colitis, wherein the active agent is selected from the group consisting of aminosalicylates, drugs. containing 5-aminosalicylic acid (5-ASA), corticosteroids, immunomodulators, cyclosporin A, TNF alpha, thiazoldinediones and glitazones. A drug delivery system according to any one of claims 1 to 6 and 9 for the treatment of a disorder such as colon cancer, wherein the active agent is selected from the group consisting of antiproliferative agents, modifying agents or DNA repair1 DNA synthesis inhibitors, DNA / RNA transcription regulators, RNA process inhibitors, agents that affect protein expression, synthesis and stability, agents that affect protein localization or its ability to exercise protein. physiological action, agents that interfere with protein-protein or protein-nucleic acid interactions, agents that act through interfering RNA, receptor binding molecules of any chemical nature (including small molecules and antibodies), labeled toxins, enzyme activators , enzyme inhibitors, gene regulators, HSP 90 inhibitors, molecules that interfere with microtubules or other cytokine components. skeletal or cellular adhesion and motility, phototherapy agents, and adjuncts to therapy. A drug delivery system according to any one of claims 1 to 6 and 9 for the treatment of disorder such as irritable bowel syndrome or constipation, wherein the active agent is selected from the group consisting of stimulant laxatives. , osmotic laxatives, stool softeners, bulking agents, tegaserod, and anticholinergic medications. A drug delivery system according to any one of claims 1 to 5 and 9, wherein the system is used as a diagnostic agent and the encapsulated agent is a diagnostic agent. A drug delivery system according to claim 17, wherein the diagnostic agent is selected from the group consisting of radiolabelled compounds, radiopaque compounds, and gases.