WO2020243393A1

WO2020243393A1 - Design of a single oral delivery system containing a monoclonal antibody for the simultaneous treatment of chron's disease and ulcerative colitis

Info

Publication number: WO2020243393A1
Application number: PCT/US2020/035071
Authority: WO
Inventors: George A. Digenis; Mahendra K. SREERAMOJU
Original assignee: Vta Labs, Llc
Priority date: 2019-05-30
Filing date: 2020-05-29
Publication date: 2020-12-03

Abstract

An oral delivery system comprising an antibody, enteric coated first minitabs, enteric coated second minitabs, and a gelatin capsule. The enteric coating on the minitabs comprise a pH-dependent enteric coating, a time-dependent enteric coating, or combination thereof. The enteric coating on the first minitabs are different from the enteric coating on the second minitabs. The antibody is within either the first, second, or both minitabs. All the minitabs are within the gelatin capsule. The oral delivery system is useful for treating Crohn's Disease, Ulcerative Colitis, or both.

Description

DESIGN OF A SINGLE ORAL DELIVERY SYSTEM

CONTAINING A MONOCLONAL ANTIBODY FOR THE SIMULTANEOUS TREATMENT OF CROHN’S

DISEASE AND ULCERATIVE COLITIS

CROSS REFERENCE TO RELATED APPLICATIONS

_[oooi_] The present application hereby claims the benefit of the provisional patent application of the same title, Serial No. 62/854,454, filed on May 30, 2019, the disclosures of which is herein incorporated by reference in its entirety.

BACKGROUND

_[0002_] Crohn’s disease and ulcerative colitis can be treated with a recombinant transgenically-expressed immunoglobulin G, subclass 1 (Ig Gl) monoclonal antibody with the amino acid sequence (aglycon portion) of adalimumab, and with a different sialyation and glycosylation; the active pharmaceutical ingredient (API). It differs from adalimumab in its polysaccharide (glycon portion) glycosylation. This active ingredient is an anti-tumor necrosis factor alpha (anti-TNFa) Ig Gl monoclonal antibody (mAb) obtained from the milk of transgenic goats (U.S. Patent No. 7,939,317). The research name of this glycoprotein was assigned to be VTA17.

_[0003] The oral site- specific delivery of a protein (including a monoclonal antibody) into the various segments of the gastrointestinal (Gl) tract is a challenge in the formulation development field since the protein must survive the proteolytic activity of two enzymes: pepsin and pancreatin. Pepsin operates in the stomach under acidic conditions (around pH 1.2), while pancreatin lyses proteins in the small intestines in a neutral environment (around pH 7.2). The proteolytic enzymatic destruction of pepsin can be circumvented by proper enteric coating of an oral dosage form.

_[0004] Pancreatin is the dominant proteolytic enzyme in the upper small intestine and its concentration can reach up to 0.9 mg/mL under fed conditions. Another major challenge associated with development of oral dosage form for antibodies (intended to deliver in intestine or colon) is preserving their integrity against enzymatic proteolysis by pancreatin. Fiterature indicates (Vipul Yadav et al.‘Gastrointestinal stability of therapeutic anti-TNF a IgGl monoclonal antibodies’ Int. J. Pharmaceutics, (2016), Vol. 502, 181) that monoclonal antibodies such as adalimumab and infliximab degrade to 90% within 30 minutes in small intestinal fluids with pancreatin. Hence it is of paramount importance to design the oral dosage form to balance the release of the active mAb and its protection against pancreatin.

BRIEF SUMMARY

_[0005] An oral delivery system comprising an antibody, enteric coated first minitabs, enteric coated second minitabs, and a gelatin capsule. The enteric coating on the minitabs comprise a pH-dependent enteric coating, a time-dependent enteric coating, or combination thereof. The enteric coating on the first minitabs are different from the enteric coating on the second minitabs. The antibody is within either the first, second, or both minitabs. All the minitabs are within the gelatin capsule. The oral delivery system is useful for treating Crohn’s Disease, Ulcerative Colitis, or both.

_[0006_] These and other objects and advantages shall be made apparent from the accompanying drawings and the description thereof.

BRIEF DESCRIPTION OF THE FIGURES

_[0007] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments, and together with the general description given above, and the detailed description of the embodiments given below, serve to explain the principles of the present disclosure.

_[0008_] Figure 1 shows release and survival profiles of VTA 17 from Intestinal Coatings 1&2 in simulated human intestinal fluids for up to 5 hours. Intestinal Coating- 1 delivers maximum dosage within two hours while Intestinal Coating-2 releases the active slowly over the period of 4 hours.

_[0009] Figure 2 shows release and survival profiles of VTA 17 from Intestinal Coating- 1 in simulated human intestinal fluids with and without pancreatin (PN). _[ooio_] Figure 3 shows release and survival profiles of VTA 17 from Intestinal Coating-2 in simulated human intestinal fluids with and without pancreatin (PN).

_[ooii_] Figure 4 shows release and survival profiles of VTA 17 from Colonic Coatings 1&2 in simulated human colonic fluids. Colonic coating- 1 with a lower percentage of time-dependent polymer undercoat releases higher amounts of the antibody than that of colonic coating-2.

_[0012_] Figure 5 shows release and survival profiles of VTA 17 from Colonic Coatings 1&2 (encased in 00 hard gelatin capsule) in gastric fluids (without pepsin), simulated human intestinal fluids (without pancreatin) and simulated human colonic fluids.

DETAILED DESCRIPTION

_[0013] An oral delivery system comprises an antibody, enteric coated first minitabs, enteric coated second minitabs, and a gelatin capsule. The enteric coating on the minitabs comprises a pH-dependent enteric coating, a time-dependent enteric coating, or combination thereof. The enteric coating on the first minitabs is different from the enteric coating on the second minitabs. The antibody is within either the first, second, or both minitabs. All the minitabs are within the gelatin capsule.

_[0014] The oral delivery system is formulated to deliver an antibody, such as VTA 17, specifically into the small intestine and the colon. The hard gelatin capsule encases multiparticulates in the form of minitabs. One portion of the minitabs was designed to deliver the antibody to the earlier portion of small intestine, for the treatment of Crohn’s Disease (CD), and the other was specifically coated to deliver additional antibody to the colon for the treatment of Ulcerative Colitis (UC).

_[0015] Examples of monoclonal antibodies include, but are not limited to VTA- 17, trastuzumab, adalimumab, bevacizumab, or combinations thereof. In some embodiments, the monoclonal antibody is VTA- 17, a recombinant transgenically expressed

immunoglobulin G (subclass 1 Ig Gl) monoclonal antibody obtained from the milk of transgenic goats (VTA17) (US Patent 7,939,317). _[0016_] The antibody may be transformed from a semi-solid in solution to a free-flowing solid state by an evaporative solidification process as described in PCT/US20/26092, incorporated by reference in its entirety. The application teaches that the produced powder exhibits a well-defined melting point with improved fluidity which enables easy mixing with various inactive ingredients.

Compression of Core minitabs

_[0017] The antibody may be mixed with a variety of excipients. The mixture of the antibody and excipients can be compressed into minitabs (<3mm) using compression pressure ranging from 3.5 to 10.5 kbar. Examples of excipients include stabilizers, such as carboxymethyl dextran (CMD), hydroxy propyl beta cyclodextrin (HPBCD), separately or in combination and basic amino acids, such as arginine HC1 and Histidine HC1 in combination with HPBCD. Use of these excipients reduces the aggregation of VTA17 (mAb) during compression and destruction by the proteolytic action of the enzyme pancreatin. Examples of additional optional excipients include, but are not limited to, binders, glidants, and lubricants.

_[0018_] The core of the minitabs are formed by mixing the antibody with other inactive ingredients then compressing the blend into minitabs. In some embodiments, the ingredients are all powders. In some embodiments, a powder formulation is prepared by blending solid VTA 17 with other inactive ingredients that are GRAS (generally regarded as safe) materials. Examples of inactive ingredients include, but are not limited to, binders such as microcrystalline cellulose (MCC), hydroxypropyl methyl cellulose (HPMC); glidants, such as silicon dioxide; and lubricants, such as magnesium stearate (see table 1).

_[0019] Aggregation (which results often in denaturation) of proteins as a function of pressure is a commonly encountered problem. Back in 1914, it was reported that a pressure of 7 kbar was able to denature proteins of egg white (Mozhaev, V. V.;

Heremans, K.; Frank, J.; Masson, P.; Balny, C., High pressure effects on protein structure and function. Proteins: Structure, Function, and Bioinformatics 1996, 24 (1), 81-91. Bridgman, P. W., The Coagulation of Albumin by Pressure. Journal of Biological Chemistry 1914, 19, 511-512.). Application of high pressure induces either local or global changes in the protein structure and finally may lead to denaturation. While the pressure of 1-2 kbar is sufficient to cause dissociation of oligomeric and multiprotein complexes, denaturation of monomeric proteins is induced at a pressure range of 4-8 kbar (J L Silva, a.; Weber, G., Pressure Stability of Proteins. Annual Review of Physical Chemistry 1993, 44 (1), 89-113. Heremans, K., High Pressure Effects on Proteins and other Biomolecules. Annual Review of Biophysics and Bioengineering 1982, 11 (1), 1- 21.).

_[0020_] In some embodiments, a powder comprises one or more monoclonal antibody, one or more cyclodextrin, and a compound selected from carboxymethyl dextran (CMD), one or more basic amino acid, or both. In some embodiments, the powder comprises about 20% to about 40% of one or more monoclonal antibody, about 35% to about 70% of one or more cyclodextrin, and about 35% to about 70% CMD. In some embodiments, the powder comprises about 20% to about 40% of one or more monoclonal antibody, about 35% to about 70% HPBCD, and about 35% to about 70% CMD. In some embodiments, the powder comprises about 20% to about 40% of one or more monoclonal antibody, about 45% to about 70% of one or more cyclodextrin, and about 15% to about 25% of one or more basic amino acid. In some embodiments, the powder comprises about 20% to about 40% of one or more monoclonal antibody, about 45% to about 70% HPBCD, and about 15% to about 25% of one or more basic amino acid.

_[0021_] In some embodiments, a powder comprises one or more monoclonal antibody, one or more cyclodextrin, carboxymethyl dextran (CMD), and one or more basic amino acid.

_[0022_] Cyclodextrins are a class of oligosaccharide macromolecules with a shape of a hollow truncated structure with hydrophilic exterior and hydrophobic interior. Examples of cyclodextrins include, but are not limited to, 2-hydroxy propyl beta cyclodextrin (HPBCD) and sulfobutylether beta cyclodextrin (SBECD). In some embodiments, the cyclodextrin is 2-hydroxy propyl beta cyclodextrin (HPBCD). In some embodiments, the cyclodextrin is sulfobutylether beta cyclodextrin (SBECD). _[0023] Carboxymethyl dextran (CMD) is a linear polymer with a (l-6)-linked glucose chains with low percentage (2-5%) of a (1-3) branches. CMDs are polyanionic in character due to the presence of about 5% negatively charged carboxyl groups (Gekko, K.; Noguchi, H., Selective interaction of calcium and magnesium ions with ionic dextran derivatives. Carbohydrate Research 1979, 69 (1), 323-326.). The molecular weights for the CMD range from about 40kDa to about 500 kDa.

Coating of minitabs

_[0024] The minitabs are coated with one or more enteric coatings. Eudragit polymer systems, based on poly(meth)acrylate chemistry provides a versatile platform to design oral drug delivery systems. Depending upon the functional groups used in the Eudragit polymer, formulations can be precisely tuned for the type of drug release such as immediate, delayed or sustained. The coatings may be pH-dependent enteric coatings, time-dependent enteric coatings, or both. A pH-dependent enteric coating is one that dissolves at a pH greater than 5.5. Examples of polymers used in pH-dependent coatings include, but are not limited to, Eudragit L100, L12.5, L100-55, and L30D 55, which dissolve at pH greater than 5.5, while the pH-dependent polymers such as Eudragit S100, S12.5, FS 30D dissolve at pH above 7.0. pH-dependent Eudragit polymers are anionic copolymers (based on methacrylic acid-methyl methacrylate or methacrylic acid-ethyl acrylate) with pH-dependent solubility character that offer enteric coatings that protect the active substance from gastric fluids in the stomach’s acidic environment (pH 1-2) until it reaches intestine or colon.

_[0025] Time-dependent Eudragit polymers are insoluble polymers with pH-independent swelling character that offer sustained and time-controlled release of active substances.

_[0026_] A time-dependent enteric coating is one that with a pH-independent swelling character that provides sustained and time-controlled release of active substances. In some embodiments, the polymers used in these time-dependent enteric coatings are hydrophobic. Over time they expand creating pores in the coating allowing a sustained release of the drug substance. Examples of polymers used in time-dependent coatings include, but are not limited to, Eudragit RS PO, RL PO, RS 100, RL 100, RS 30 D, RS 12.5, RL 30 D and RL 12.5. The coating may contain additional inactive ingredients such as opacifiers, glidants, and plasticizers, including organic solvents such as ethanol and isopropyl alcohol.

_[0027] The difference in the properties of pH-dependent enteric coatings and time- dependent enteric coatings can be seen in figure 1. The Intestinal Coating- 1 that is coated exclusively with pH-dependent polymer (L100) open up immediately in intestinal fluids (pH > 5.5) and release almost 80% of active substance in less than 1 hour. Whereas Intestinal Coating-2 that has pH-dependent polymer coating (LI 00) with an undercoat of time-dependent polymer (RS PO) didn’t release the active immediately but shows a sustained release over the time period of 4 hours with maximum reaching after 2 hours.

_[0028_] Examples of pH-dependent coatings include those that comprise one or more polymer selected from Eudragit L-100, L12.5, L100-55, L30D 55, S100, S12.5, and FS 30D. Examples of time-dependent coatings include those that comprise one or more polymer selected from Eudragit RS PO, RL PO, RS 100, RL 100, RS 30 D, RS 12.5, RL 30 D and RL 12.5; such as RS PO, RL PO, RS 100, and RL 100. In some embodiments, the coating comprises up to 5% time-dependent polymer, such as 0% to about 5%, about 0.1% to about 5%, about 0.1% to about 3%, about 0.1% to about 2.5%, about 0.1% to about 2%, about 0.1% to about 1%, about 0.5% to about 3%, about 0.5% to about 2%, about 0.5% to about 1%, about 1% to about 3%, about 1% to about 2%, about 5.5%, about 2.4%, and about 1.8%. In some embodiments, the coating comprises about 1% to about 20% pH-dependent polymer, such as about 1% to about 15%, about 1% to about 12%, about 1% to about 10%, about 5% to about 20%, about 5% to about 15%, about 5% to about 12%, about 5% to about 10%, about 7% to about 20%, about 7% to about 15%, about 7% to about 12%, about 7% to about 10%, about 12.6%, about 11%, and about 10%. In some embodiments, the coating comprises both time-dependent polymers and pH-dependent polymers. The amounts may be any combination of the above.

_[0029] The coating may be formed from a solution. In some embodiments, the coating solution comprises Eudragit polymer (pH-dependent or time-dependent), triethyl citrate as a plasticizer, titanium dioxide as an opacifier, and magnesium stearate as glidant; all dissolved or well suspended in a solvent mixture of isopropanol/ethanol/water in (90/5/5) ratio.

_[0030] The minitab coating was performed on Freund- Vector VFC-LAB Micro FLO- Coater® Bench top fluid bed system with a top-spray coating system. High-pressure nitrogen gas was used to fluidize the minitabs in the coating chamber. The minitabs were fluidized initially for 5 minutes at an inlet temperature of 35 °C to dry the minitabs before coating. The coating solution was transferred into the chamber using a peristatic pump and sprayed on the fluidized minitabs using top-spray method. Percent weight gain was recorded after several cycles of coating.

_[0031] The oral delivery system comprises two types of minitabs, the first minitabs and the second minitabs. The enteric coatings on the first and second minitabs are different. The different enteric coatings allow the antibody to be released at different locations of the intestinal tract, such as the small intestine and the colon. In one embodiment, the enterically coated minitabs are intended to release antibody VTA17.

_[0032] In one embodiment, an enteric-coated minitabs released 78% of intact VTA17 after 60 minutes of exposure to simulated human intestinal fluid containing maximum concentration of pancreatin (0.95 mg/mL at 37°C) (Figure 1, table 5). Figure 1 and Table 5 show the release profile of both Intestinal Coating- 1 and Intestinal Coating-2.

_[0033] In one embodiment, an enteric-coated minitabs released 50% of intact VTA17 after 2 hours of exposure to simulated human colonic fluid. The appropriately coated minitabs released 50% of intact VTA17 in 2 hours and 63% in 4 hours of incubation.

This is the colonic incubation where there is no proteolytic degradation. Hence with time, the amount of release will increase.

_[0034] The minitabs are within a gelatin capsule. In some embodiments, the gelatin capsule is a hard gelatin capsule of size Ό’ or Ό0’. In some embodiments, there are from about 70 to about 75 minitabs within the gelatin capsule. The amount of first and second minitabs in the gelatin capsule will determine the amount of antibody delivered to different parts of the intestinal tract. In some embodiments, the ratio of amounts of first minitabs to second minitabs ranges from about 1:5 to about 5:1. In some embodiments, the amount of antibody in one gelatin capsule ranges from about 30 mg to about 80 mg.

_[0035] The gelatin capsule will be formed by selecting the number of intestinal and colonic minitabs, filling the body of the capsule with them, and closing the cap.

_[0036] The oral delivery system may be used to treat patients with Crohn’s Disease (suffering from the existence of lesions in the small intestines), Ulcerative Colitis (lesions in the colon), or both.

Treatment of Crohn’s disease

_[0037] Various proportions of time-dependent (Eudragit RS PO) and pH-dependent (Eudragit L-100) polymers may be used to coat the minitabs, which are intended to be delivered in the small intestine. An example of two coatings are Intestinal Coatings 1 & 2. Table 3 shows the coating composition of these two coatings. Intestinal Coating- 1 was coated in such a way to release major portion of VTA17 at the earlier part of small intestine, such as jejunum. Intestinal Coating-2 was coated to deliver its contents at the later part of small intestine such as the ileum. To achieve a homogeneous and uniform coating with prescribed thickness, a fluidized-bed film coating process (Wurster) was utilized.

Table 3: Intestinal Coatings with percent coating on surface of minitabs

% Coating

Time-Dependent pH-Dependent

Coating (Eudragit RS PO) (Eudragit L-100)

Intestinal Coating -1 0 10

Intestinal Coating -2 1.8 10

_[0038] Figure 1 shows the in vitro release and survival of VTA17 from Intestinal Coatings 1&2 in simulated human intestinal fluid (containing pancreatin at maximum concentration of 0.9 mg/mL) at 37 °C and stirred at 55 RPM. Simulated intestinal fluids were prepared by dissolving 12.5 g of NaHC03, 6 g of dehydrated bile extract and 0.9 g of pancreatin in one liter of de-ionized (DI) water (composition shown in Table 4). The pH of the resulting solution was adjusted to 6.8 using IN HC1. The concentration of pancreatin (0.9 mg/mL) in simulated intestinal fluids used for the present study was at its maximum concentration that usually is attained only under fed conditions. Thus, the release studies of these coatings were conducted in intestinal fluids with much higher pancreatin concentration than normal.

Table 4: Composition of simulated human intestinal fluid used for in vitro release studies

S. No. Ingredient Amount/ 1 liter

1 Sodium bicarbonate (NaHCOB) 12.5 g

2 Dehydrated bile extract 6 g

3 Pancreatin 0.9

_[0039] As it can be seen from Figure 1 and Table 5, Intestinal Coatings 1 & 2 were used for the in vitro release studies. Intestinal Coating- 1 was coated exclusively with pH- dependent polymer of Eudragit L-100 (Table 3). Intestinal Coating- 1 released its maximum content in about one hour after its incubation in simulated human intestinal fluids. Interestingly, about 78% of VTA17 from Intestinal Coating -1 was released and survived after one hour of proteolytic action by pancreatin; and about 50% of VTA17 released and survived after two hours of proteolytic action by pancreatin (Figure 1 and Table 5). Intestinal Coating-2 was coated with 10% of Eudragit L-100 with an additional undercoat of 1.8% of time-dependent Eudragit RS PO (see Table 3). The VTA17 released and survived from Intestinal Coating-2 in one hour was about 10% and about 50% after 2 hours of incubation in simulated human intestinal fluid with maximum pancreatin concentration. Thus, Intestinal Coating-2 appeared to release its contents at slower rate than Intestinal Coating- 1 due to its extra undercoat with 1.8% of time-dependent polymer, Eudragit RS PO in addition to an enteric coating layer of 10% Eudragit L-100.

% Release & Survived

Intestinal Coating-2

Intestinal Coating-1 (1.8% Eudragit RS PO

S. No. Time (h) (10% Eudragit L100) & 10% Eudragit L100)

1 0.3 39.4 1.2

2 0.6 71.8 2.6

3 1.0 76.5 10.6

4 2.0 47.6 42.5

5 3.0 25.9 35.4

6 4.0 15.0 30.0

_[0040] Also, after 4 hours of intestinal incubation with pancreatin at its maximum concentration, about 15% of VTA17 from Intestinal Coating-1 and 30% of VTA17 from Intestinal Coating-2 survived and would have survived a typical transit time through the intestine into the colon.

_[0041] The release and survival profile of both Intestinal Coatings 1&2 were tested in simulated human intestinal fluids without pancreatin. This study was conducted to ascertain that the coated minitabs release maximum of their VTA17 content when dispersed in intestinal fluids. Figure 2 and table 6 show a comparison in the release and survival profiles of VTA 17 from the Intestinal Coating -1 in simulated human intestinal fluids with and without pancreatin. Intestinal Coating- 1, in the absence of pancreatin, released its maximum content (about 95%) of VTA17 in less than 1 hour. In 1 hour, in the presence of pancreatin, the amount of VTA17 released and survived was about 76%. From this data, it can be concluded that about 19% of VTA17 was proteolyzed by pancreatin in 1 hour. Table 6: Release and survival profile of VTA / 7 from Intestinal Coating-1 in simulated _ human intestinal fluids with and without pancreatin (PN) _

% Released & Survived (Intestinal Coating -

1)

(10% Eudragit L100)

S. No. Time (h) With Pancreatin Without Pancreatin

1 0.3 39.4 65.5

2 0.6 71.8 89.5

3 1.0 76.5 95.5

4 2.0 47.6 97.5

5 3.0 25.9 98.0

6 4.0 15.0 98.9

_[0042] The formulation (Intestinal Coating- 1) designed for intestinal delivery is efficient enough not only in releasing its maximum dose in one hour but can also protect VTA17 from pancreatin to a considerable extent so that about 76% of VTA17 was released and survived after one hour and about 50% released and survived after two hours. Similarly, Intestinal Coating-2 released its maximum of VTA 17 (about 95%) in 4 hours in simulated human intestinal fluids without pancreatin (Figure 3 and Table 7). As expected, with an additional undercoat with 1.8% of time-dependent polymer (Eudragit RS PO), the release was delayed. The release profile of Intestinal Coating-2 indicates that it is a right choice to deliver the contents at the latter part of intestine such as ileum.

Table 7: Release and survival profile of VTA / 7 from Intestinal Coating-2 in simulated _ human intestinal fluids with and without pancreatin (PN) _

% Released & Survived (Intestinal Coating -

2)

(1.8% Eudragit RS PO & 10% Eudragit L100)

S. No. Time (h) With Pancreatin Without Pancreatin

1 0.3 1.2 0

2 0.6 2.6 1.4

3 1.0 10.6 7.0

4 2.0 42.5 45.5

5 3.0 35.4 84.8

6 4.0 30.0 94.5

Treatment of Ulcerative Colitis

_[0043] Various proportions of time-dependent (Eudragit RS PO) and pH-dependent (Eudragit L100) polymers may be used to coat the minitabs, which are intended to be delivered in the colon. Examples of two coatings are Colonic Coatings 1 & 2. Table 8 shows the composition of these two coatings. Both Colonic Coatings 1 & 2 had time- dependent polymer (Eudragit RS PO) layer first (5.5% for Colonic Coating- 1 and 2.4% for Colonic Coating-2), followed by pH-dependent polymer (Eudragit S100) layer (11% for Colonic Coating- 1 and 12.6% for Colonic Coating-2) as an outer layer (Table 8).

Table 8: Colonic Coatings with percent coating on the surface of minitabs

% Coating

Time-Dependent pH-Dependent

Coating (Eudragit RS PO) (Eudragit S100)

Colonic Coating -1 5.5 11

Colonic Coating -2 2.4 12.6

_[0044] The release and survival profile of Colonic Coatings 1 & 2 were tested in simulated human colonic fluids. The materials used to prepare simulated colonic fluids (FeSSCoF) are procured from Biorelevant. In the first step, tris/maleate buffer was prepared by dissolving 0.66 g of NaOH, 3.5 g of maleic acid and 3.69 g of tris base in 1 liter of water and the pH was adjusted to 6.0 using either 1 N NaOH or 1 N HC1. In the second step, about 0.74 g of FeSSCoF powder was added to 1 liter of already prepared buffer and dissolved. The composition of simulated human colonic fluids is shown in Table 9.

Table 9: Composition of simulated human colonic fluid used for in vitro release studies

Amount

S. No. Ingredient (g/Liter)

1 Sodium hydroxide 0.66

2 Maleic acid 3.5

3 Tris base 3.69

4 FeSSCoF 0.74

_[0045] As can be seen from Figure 4 and Table 10, both Colonic Coatings exhibited a sustained release profile for VTA17 as a function of time-dependent polymer (Eudragit RS PO) coating for up to 24 hours. Colonic Coating- 1, which is coated with 11% of pH- dependent polymer (Eudragit S100) and 5.5% of time-dependent polymer (Eudragit RS PO) had about 47% of VTA17 released and survived in 24 hours. Colonic Coating-2, that had about 12.5% of enteric coating, but lesser percent (2.4%) of time dependent polymer coating had a greater amount of VTA17 (63%) released and survived in 24 hours.

% Released & Survived

Colonic Coating-2

Colonic Coating-1 (2.4% Eudragit RS PO

(5.5% Eudragit RS PO & 12.6% Eudragit

S. No. Time (h) & 11% Eudragit S100) S100)

1 1.0 1.3 1.2

2 8.5 23.7 49.5

3 20 43.4 61.8

4 24 47.8 63.3 _[0046] In another separate study, a 00-sized hard gelatin capsule encasing Colonic Coatings was incubated in gastric fluid (without pepsin) for 1.5 hours at 37 °C while stirring at 55 RPM. After 1.5 hours of incubation in gastric fluids, the liberated minitabs from the gelatin capsule were transferred into simulated human intestinal fluids (without pancreatin), incubated for 3 hours at 37 °C while stirring at 55 RPM; subsequently these minitabs were transferred into simulated human colonic fluids and incubated for 24 hours at 37 °C while stirring at 55 RPM. The release and survival of VTA17 at various time intervals during this study were shown in Figure 5 and Table 11.

Table 11: Release and survival profiles of Colonic Coatings 1 & 2 ( encased in 00 hard gelatin capsule ) in gastric fluids (without pepsin), simulated intestinal fluids (without pancreatin) followed by simulated human colonic fluids.

% Released & Survived

Colonic Coating-2

Colonic Coating-1 (2.4% Eudragit RS PO

Time (5.5% Eudragit RS PO & 12.6% Eudragit

System (h) & 11% Eudragit S100) S100)

Stomach 0 0 0

(without Pepsin) 1.5 0 0

Small Intestine 2.5 0.9 0.7

(Without

Pancreatin) 4.5 6.3 17.0

5 .5 9.9 28.9

6.5 15.0 38.2

7.5 20.4 46.0

9 25.0 51.1

12 32.2 57.3

Simulated Human 24 46.0 61.3

Colonic Fluid 27 47.4 62.3

_[0047] This study showed that the Colonic Coatings had enough thickness of enteric coating that they remained intact during incubation in gastric fluids. However, these Colonic Coatings did release a fraction of dose in simulated human intestinal fluids. As shown in Figure 5 and Table 11, Colonic Coating-1 release about 6% while Colonic Coating-2 release about 17% of VTA17 in intestinal fluids after 3 hours of incubation.

_[0048] Literature indicates (Vipul Yadav et al.‘Gastrointestinal stability of therapeutic anti-TNF a lgGl monoclonal antibodies’ Int. J. Pharmaceutics, (2016), Vol. 502, 181) that other monoclonal antibodies (like adalimumab and infliximab) degrade to 90% within 30 minutes in small intestinal fluids. In contrast, the formulation described has a 78% survival of VTA17 for a period of 1 hour and 50% survival for a period of 2 hours in simulated human intestinal fluid at 37°C. It also survived for at least 12 hours in the presence of simulated human colonic fluids. The in-vitro release and survival profiles of both Cntestinal and Colonic Coatings indicate that the developed coatings for the treatment of Crohn’s disease and ulcerative colitis site- specifically deliver desirable amounts of VTA17 in a stable form without much altering the integrity of antibody, VTA17.

_[0049] A combination of Intestinal Coating - 1 and Colonic Coating -2 in desirable proportion (loaded in 00 or 0 sized capsule) may be an appropriate combination to simultaneously treat patients with Crohn’s disease (CD) and ulcerative colitis (UC).

_[0050] While the present disclosure has illustrated by description several embodiments and while the illustrative embodiments have been described in considerable detail, it is not the intention of the applicant to restrict or in any way limit the scope of the appended claims to such detail. Additional advantages and modifications may readily appear to those skilled in the art. Furthermore, features from separate lists can be combined; and features from the examples can be generalized to the whole disclosure.

EXAMPLES

Example 1: Formation of the core of the minitabs

_[0051] The powder blends in Table 1 were thoroughly mixed, except the magnesium stearate, which was added and mixed later, just before compression. These blends were compressed into minitabs of 2 mm diameter at pressure ranging from 3.5 to 10.5 kbar. The various ingredient compositions used in minitabs compression and their function are listed in Table 1.

_ Table 1: Composition o f minitab matrix _

Composition

Composition 1 2 Ingredient

S. No. Ingredient % (w/w) % (w/w) Function

1 mAb (VTA17) 11.5 12.1 Active Ingredient

2 Co-Proteins 8.3 0

3 Sodium Acetate 0.9 1 Buffer Component

2-Hydroxypropyl b-

4 cyclodextrin (HPBCD) 11.5 - 28 12 - 30 Stabilizer

Carboxymethyl dextran

5 (CMD) 11.5 - 28 0 Stabilizer

Basic amino acid

6 (Arginine/Histidine) 0 12 - 24 Stabilizer Microcrystalline Cellulose

7 (MCC) 32 36 Binder

Hydroxypropyl methyl

8 cellulose (HPMC) 2.3 2.7 Binder

9 Silicon dioxide 1.3 1.5 Lubricant

10 Magnesium stearate 3.6 4.1 Glidant

Composition 1 is for 60% pure VTA17 and composition 2 is for 99% pure VTA17

Example 2: Coating minitabs

_[0052] The minitab coating was performed on Freund- Vector VFC-LAB Micro FLO- Coater® Bench top fluid bed system with a top-spray coating system. High-pressure nitrogen gas was used to fluidize the minitabs in the coating chamber. The minitabs were fluidized initially for 5 minutes at an inlet temperature of 35 °C to dry the minitabs before coating. The coating solution was transferred into the chamber using a peristatic pump and sprayed on the fluidized minitabs using top-spray method. Percent weight gain was recorded after several cycles of coating.

_[0053] The composition of ingredients in the coating solution is shown in Table 2. Table 2: Composition of coating solution

S. No. Ingredient % (w/V)

1 Eudragit Polymer 7 - 10%

2 Triethyl citrate 0.75

3 Titanium dioxide 0.25

4 Magnesium stearate 0.25

Example 3: Experimental for Crohn’s disease minitabs and testing

[_0054] The enterically coated minitabs (Intestinal Coating- 1) which are intended to release their antibody, VTA17, in simulated human intestinal fluid containing maximum concentration of pancreatin (0.95 mg/mL at 37°C) were found to release and survive 78% of intact VTA17 in 60 minutes (Figure 1, table 5). The appropriately coated minitabs released 50% of intact VTA17 in 2 hours and 63% in 4 hours of incubation (Figure 2). This is the colonic incubation where there is no proteolytic degradation. Hence with time, the amount of release will increase.

_[0055] The coating was performed on Freund- Vector VFC-LAB Micro FLO-Coater® Bench top fluid bed system with a top-spray coating system. High-pressure nitrogen gas was used to fluidize the minitabs in the coating chamber. The minitabs were fluidized initially for 5 minutes at an inlet temperature of 35 °C to dry the minitabs before coating. The coating solution was transferred into the chamber using a peristatic pump and sprayed on the fluidized minitabs using top-spray method. Percent weight gain was recorded after several cycles of coating.

_[0056] Size exclusion chromatography (SEC) was used to find out the amounts of antibody released and survived at various time-intervals of intestinal and colonic incubation. The aliquots of incubated samples were collected, dissolved in phosphate buffer (PBS), filtered using 0.2 pm PVDF membrane filters and analyzed using HPLC equipped with SEC column. An Acquity UPLC protein BEH200 SEC column (4.6 x 150 mm, 1.7 pm) was used as stationary phase and phosphate buffer as mobile phase was used for the analysis.

Claims

CLAIMS What is claimed is:

1. An oral delivery system comprising an antibody, enteric coated first minitabs, enteric coated second minitabs, and a gelatin capsule;

wherein the enteric coating on the minitabs comprise a pH-dependent enteric coating, a time-dependent enteric coating, or combination thereof;

wherein the enteric coating on the first minitabs are different from the enteric coating on the second minitabs;

wherein the antibody is within either the first, second, or both minitabs; and

wherein all the minitabs are within the gelatin capsule.

2. The oral delivery system of claim 1, wherein the antibody comprises at least one of VTA17, trastuzumab, adalimumab, bevacizumab, or combinations thereof.

3. The oral delivery system of any one of claims 1 to 2, wherein the pH-dependent coating comprises one or more polymer selected from Eudragit L100, L12.5, L100-55, L30D 55, S100, S12.5, and FS 30D.

4. The oral delivery system of any one of claims 1 to 3, wherein the time-dependent coating comprises one or more polymer selected from Eudragit RS PO, RL PO, RS 100, and RL 100.

5. The oral delivery system of any one of claims 1 to 4, wherein the minitabs

comprise a mixture of one or more antibody, one or more cyclodextrin, and a compound selected from carboxymethyl dextran (CMD), one or more basic amino acid, or both.

6. The oral delivery system of any one of claims 1 to 5, wherein the pH-dependent enteric coating comprises Eudragit S100 and the time-dependent enteric coating comprises Eudragit RS PO.

7. The oral delivery system of any one of claims 1 to 6, wherein the ratio of first minitabs to second minitabs ranges from about 1:5 to about 5:1.

8. The oral delivery system of any one of claims 1 to 7, wherein the amount of antibody ranges from about 30 mg to about 80 mg.

9. The oral delivery system of any one of claims 1 to 8, wherein the gelatin capsule is a hard gelatin capsule of size O’ or Ό0’ .

10. A method for treating Crohn’s Disease, Ulcerative Colitis, or both, comprising administering the oral delivery system of any one of claims 1 to 9 to a subject in need of said treatment.

11. The method of claim 10, wherein the antibody comprises at least one of VTA17, trastuzumab, adalimumab, bevacizumab, or combinations thereof.

12. The method of any one of claims 10 to 11, wherein the minitabs comprise a mixture of one or more antibody, one or more cyclodextrin, and a compound selected from carboxymethyl dextran (CMD), one or more basic amino acid, or both.

13. The method of any one of claims 10 to 12, wherein antibody is released from the first minitabs in the small intestine and from the second minitabs in the colon.

14. The method of any one of claims 10 to 13, wherein the pH-dependent coating comprises one or more polymer selected from Eudragit L100, L12.5, L100-55, L30D 55, S100, S12.5, and FS 30D.

15. The method of any one of claims 10 to 14, wherein the time-dependent coating comprises one or more polymer selected from Eudragit RS PO, RL PO, RS 100, and RL 100.

16. The method of any one of claims 10 to 15, wherein the pH-dependent enteric coating comprises Eudragit S100 and the time-dependent enteric coating comprises Eudragit RS PO.

17. The method of any one of claims 10 to 16, wherein the ratio of first minitabs to second minitabs ranges from about 1:5 to about 5:1.

18. The method of any one of claims 10 to 17, wherein the amount of antibody ranges from about 30 mg to about 80 mg.