Cross-linked maltodextrins for oral delivery of biologically active agents

Abstract

To provide a delivery system capable of improving pharmacokinetic properties and bioavailability of insulin, for safe and efficient oral delivery of biological actives, in particular insulin.SOLUTION: The present invention provides a composition intended for oral administration of a biological active, comprising a cross-linked maltodextrin and an effective quantity of the biological active.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to the oral administration of biologically active substances, particularly insulin, and involves the use of crosslinked starch materials as a delivery system. [Background technology]

[0002] The gastrointestinal tract presents a mucosal surface that separates the internal structures from the outside world, composed of a single layer of polarized epithelial cells. Despite their delicate nature, these epithelial sheets provide a formidable barrier against the accidental and unregulated uptake of most substances to which the body is exposed through ingestion.

[0003] While protecting the body from the many toxins and pathogens it is exposed to daily, this barrier also hinders the efficient uptake of biopharmaceuticals, including peptides, proteins, and DNA- and RNA-based drugs.

[0004] This is even more challenging for the oral route. Before reaching the systemic circulation, biopharmaceuticals must retain their intact structure, integrity, and conformation through the stomach and intestine. However, biopharmaceuticals are particularly sensitive to the gastrointestinal environment, mainly due to high gastric acidity and the presence of hydrolytic enzymes.

[0005] Furthermore, the large molecular size and / or hydrophilic nature of biopharmaceuticals such as proteins prevents them from diffusing across the mucin barrier, a thick layer of mucus that coats the gastrointestinal epithelium.

[0006] As a result, parenteral routes are commonly used for the administration of biologically active substances.

[0007] However, such routes of administration have inherent drawbacks due to their invasiveness: they cause localized pain, discomfort, irritation, needle stick injuries, and risk of infection.

[0008] Among the well-known biologically active substances currently administered parenterally is insulin, which requires subcutaneous injection.

[0009] This systemic insulin administration route also has the disadvantage that only a small amount of insulin can successfully reach the liver for its own physiological action. Retention of insulin in the peripheral circulation can lead to peripheral insulin resistance and immunogenicity. Hyperinsulinemia can also occur when the injected insulin acts on the wrong target, causing hypertension, cardiovascular disease, weight gain and peripheral edema.

[0010] From the above, it follows that delivery of insulin (and more generally biologically active substances) via the oral route has several advantages over systemic administration routes, which may in particular help improve patient comfort and compliance with treatment.

[0011] Interestingly, the use of cyclodextrins crosslinked with pyromellitic dianhydride as a delivery system for oral administration of insulin has been proposed recently in patent application ITBO20120710. It has been shown that insulin loaded on such crosslinked materials can not only remain intact in the gastrointestinal tract, but also cross the membrane and enter the plasma compartment.

[0012] However, these delivery systems still needed to be improved. Oral ingestion of insulin with this delivery system causes a high peak of blood insulin after administration, which can lead to harmful hypoglycemia. Furthermore, blood insulin decreases very quickly after ingestion, which means that repeated administration is necessary to maintain blood insulin at a satisfactory level over time.

[0013] Thus, there remains an unsatisfied need for improved and safe delivery systems for the oral administration of insulin, and more generally, biologically active substances. More specifically, there remains a need for delivery systems that can improve the pharmacokinetic behavior of these biologically active substances. [Prior art documents] [Patent documents]

[0014] [Patent Document 1] ITBO20120710 Summary of the Invention [Problem to be solved by the invention]

[0015] It is therefore an object of the present invention to provide a safe and efficient system for the oral delivery of biologically active substances, particularly insulin.

[0016] In particular, it is an object of the present invention to provide a delivery system capable of improving the pharmacokinetic properties and bioavailability of orally administered biologically active substances. [Means for solving the problem]

[0017] Presentation of the invention The present inventors have successfully improved the shortcomings of the prior art techniques, in particular the crosslinked cyclodextrins of patent application ITBO20120710, by developing a novel and safe system for parenteral delivery of biologically active substances, associated with greater bioavailability and pharmacokinetic profile of the biologically active substances.

[0018] The delivery system of the present invention is characterized by the use of cross-linked maltodextrin.

[0019] The delivery system according to the invention is able to significantly increase the pharmacokinetic profile of the active protein after oral administration. This is evident from the following examples, in which the inventors have demonstrated that oral administration of insulin with the delivery system of the invention can provide a particularly longer and longer-lasting effect than that obtained with the crosslinked cyclodextrins of patent application ITBO20120710. In particular, the inventors have shown that the delivery system of the invention advantageously causes a slower release of insulin. This means that for the same amount of insulin administered, it allows insulin release over a longer period of time and can therefore be active for longer. Furthermore, the peak of insulin in the blood observed after oral administration is two times lower than that observed with the delivery system of the prior art. As a result, the delivery system of the invention is safer, since there is less risk of hypoglycemia after administration.

[0020] The delivery system of the present invention also has the advantage of being based on a starchy material, which is a bio-based material that is easily obtained and can be modified, and is well tolerated.

[0021] Summary of the Invention The present invention therefore first relates to a composition intended for the oral administration of a biologically active substance, comprising cross-linked maltodextrin and an effective amount of said biologically active substance.

[0022] The present invention also relates to a method for preparing a composition according to the invention, comprising the step of loading the crosslinked maltodextrin with an effective amount of a biologically active agent.

[0023] The present invention also relates to the use of cross-linked maltodextrins for the oral delivery of biologically active agents.

[0024] The present invention also relates to a method for treating an individual comprising oral administration of a composition according to the present invention.

[0025] In a further aspect, the present invention relates to a sustained release oral formulation, i.e. a sustained release oral formulation, comprising the composition of the present invention and suitable excipients. The oral formulation may be a pharmaceutical or veterinary product, or a product selected from the group consisting of cosmetics, food and dietary supplements. [Brief description of the drawings]

[0026] [Figure 1] Figure 1: Percentage of in vitro insulin released over time from different delivery systems at pH 1.2. [Diagram 2] Figure 2: Percentage of in vitro insulin released over time from different delivery systems at pH 6.8. [Diagram 3] Figure 3: In vivo blood insulin levels after oral administration of different insulin-loaded delivery systems to mice. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] Thus, the present invention relates to the use of cross-linked maltodextrins for the oral delivery of biologically active substances, in particular insulin.

[0028] Thus, in a first aspect, the present invention relates to a composition intended for oral administration of a biologically active substance, said composition comprising a cross-linked maltodextrin and an effective amount of said biologically active substance.

[0029] The term "maltodextrin" classically refers to a starchy material obtained by acid and / or enzymatic hydrolysis of starch. With reference to regulatory status, maltodextrins have a dextrose equivalent (DE) of 1 to 20.

[0030] Preferably, the maltodextrins useful according to the invention have a DE chosen in the range of from 5 to 20, preferably from 10 to 20, preferably from 15 to 20. This DE is for example equal to 17.

[0031] Preferably, the maltodextrins useful in the present invention are derived from starch containing 25-50% amylose, expressed as dry weight relative to the total dry weight of said starch.

[0032] This amylose content can classically be determined by one skilled in the art by potentiometric analysis of iodine, which is absorbed by amylose to form a complex.

[0033] Preferably, the maltodextrins useful in the present invention are derived from starches exhibiting an amylose content selected within the range of 25-45%, preferably 30-45%, preferably 35-40%, these percentages being expressed as the dry weight of amylose relative to the total dry weight of the starch.

[0034] It should be noted that the expression "starch" classically refers to starch isolated from any suitable plant source by any technique known to those skilled in the art. Isolated starch typically contains 3% or less impurities, said percentage being expressed as the dry weight of the impurities relative to the total dry weight of the isolated starch. These impurities typically include proteins, colloidal substances and fibrous residues. Suitable plant sources include, for example, legumes, cereals and tubers. In this regard, the starch of the present invention is preferably a legume starch, more preferably a pea starch, and even more preferably a smooth pea starch.

[0035] Preferably, the maltodextrin useful in the present invention has a weight average molecular weight selected within the range of 5,000 to 15,000 Daltons (Da), preferably 10,000 to 15,000 Da, preferably 10,000 to 14,000, for example 12,000 Da.

[0036] This weight average molecular weight can be determined by one skilled in the art, inter alia, by liquid chromatography with differential refractometric detection, preferably by using pullulan standards.

[0037] The maltodextrins useful in the present invention are obtained by hydrolysis of starch, but may have undergone other chemical and / or physical modifications, provided they do not interfere with the desired properties, particularly in terms of safety and efficiency of the final crosslinked maltodextrin. However, the maltodextrins useful in the present invention are preferably not further modified, as this is not believed to be necessary in the present invention.

[0038] Suitable maltodextrins are commercially available, for example those sold under the name KLEPTOSE® Linecaps (ROQUETTE).

[0039] The crosslinked maltodextrin useful in the present invention can be obtained (or obtained) by reacting maltodextrin with a crosslinking compound. The crosslinking compound is typically multifunctional, i.e., the crosslinking compound contains at least two reactive functional groups. Preferably, the crosslinking compound is bifunctional. Typically in the present invention, the reactive functional group has a carbon that undergoes nucleophilic attack, i.e., has a partial positive charge.

[0040] Preferably, the cross-linking compound of the present invention is selected from dianhydrides. Even more preferably, the cross-linking compound of the present invention is pyromellitic dianhydride.

[0041] Preferably, the crosslinked maltodextrin of the present invention comprises the following pattern: [ka]

[0042] wherein R1, R2, R3, and R4 are independently selected from hydrogen atoms or anhydroglucose units of maltodextrin, the number of which or said anhydroglucose units in said pattern being at least equal to 2. Preferably, the number of said anhydroglucose units is equal to 2, i.e., two of the groups R1 to R4 are hydrogen and two of the groups R1 to R4 are anhydroglucose units.

[0043] This crosslinked maltodextrin can in particular be obtained by reacting maltodextrin with pyromellitic dianhydride. Generally, when such a crosslinking compound is used, R1 and R3 are not both anhydroglucose units, and R2 and R4 are not both anhydroglucose units.

[0044] Preferably, for the cross-linking reaction, the molar ratio between the cross-linking compound and the anhydroglucose units of said maltodextrin is selected in the range of 0.1 to 10.0, more preferably 0.1 to 5.0, even more preferably 0.1 to 1.0, even more preferably 0.5 to 0.7, for example equal to 0.6, i.e. 0.6 moles of cross-linking compound are used for 1 mole of anhydroglucose units.

[0045] More specifically, the crosslinked maltodextrin of the present invention is preferably obtainable (or obtained by) a process comprising the following steps: a) preparing a solution of maltodextrin, said maltodextrin preferably being similar to those described above; b) adding at least one cross-linking compound, said cross-linking compound being preferably similar to those described above; c) Obtaining cross-linked maltodextrin.

[0046] Preferably the crosslinked maltodextrin of the present invention, especially the crosslinked maltodextrin for oral administration loaded with a biologically active agent, is in the form of particles.

[0047] In that case, the average diameter of the crosslinked maltodextrin particles useful in the present invention is selected in the range of 1 to 1000 nm. It is preferably at least 10 nm, more preferably at least 50 nm, even more preferably at least 100 nm. It is, for example, selected in the range of 100 to 1000 nm.

[0048] This average diameter is the hydrodynamic diameter, which can be determined by one of ordinary skill in the art by laser light scattering, for example, according to the procedures detailed in the Examples below.

[0049] Preferably, the polydispersity index for said average diameter is less than 1.00. This polydispersity index is generally greater than 0.05.

[0050] Preferably, the crosslinked maltodextrins of the present invention have a zeta potential of less than 0 mV, preferably less than -10 mV, or even less than -20 mV. This zeta potential is generally greater than -50 mV and greater than -40 mV.

[0051] This zeta potential can be determined by the skilled artisan by electrophoretic mobility by dynamic light scattering at a scattering angle of 90° for a crosslinked maltodextrin suspension at a temperature of 25° C. It can be determined, for example, according to the procedure detailed in the examples below.

[0052] Preferably, the loading capacity of the crosslinked maltodextrin of the present invention is at least 1%, said percentage representing the dry weight of biologically active substance relative to the total dry weight of the loaded crosslinked maltodextrin loaded with said biologically active substance.Preferably, this loading capacity is at least 5%, preferably at least 10%.This loading capacity is generally less than 30%, even less than 20%.

[0053] This loading capacity can be determined by one of skill in the art by HPLC, for example, according to the procedures detailed in the Examples below.

[0054] The crosslinked maltodextrin of the present invention may contain other compounds in its structure than the maltodextrin and crosslinking compounds useful in the present invention, as long as they do not interfere with the desired properties of said crosslinked maltodextrin, especially in terms of efficiency and safety. By "other compounds", it is understood that the inventors do not intend to include impurities, such as impurities derived from starch used as raw material for the preparation of maltodextrin. Such other compounds are classically other polymers than the maltodextrin and / or other crosslinking compounds useful in the present invention. They typically represent 10% by weight or less of the total compounds used to obtain the crosslinked maltodextrin. However, more preferably, the crosslinked maltodextrin of the present invention does not contain other compounds in its structure than the maltodextrin and crosslinking compounds useful in the present invention.

[0055] The crosslinked maltodextrins of the present invention are particularly useful for the oral administration of biologically active agents.

[0056] According to the present invention, by the definition of "biologically active" is intended any type of pharmaceutical or non-pharmaceutical biological substance that is delivered by the oral route.

[0057] The expression "biologically active" classically refers to active substances such as nucleic acid-based active substances, such as protein, DNA or RNA-based active substances, cell-based active substances, and virus-based active substances. In particular, these active ingredients are characterized by the fact that they are not bioavailable when administered orally, for example, because such routes of administration cause their degradation and / or because they are unable to cross biological barriers. In other words, these are active substances that are typically only bioavailable when administered parenterally. These active substances are typically active substances for which a systemic effect is desired. Such active ingredients, if they are ingested as such, typically (i) are degraded through the gastrointestinal tract (GIT), for example, due to the low pH of the stomach or due to enzymatic degradation, and / or (ii) are unable to cross the (GIT) barrier, for example, due to their size and / or polarity.

[0058] Preferably, the bioactive substance according to the present invention is a protein. In the context of the present invention, the term "protein" is broadly defined as commonly understood by those skilled in the art. It particularly includes proteins, regardless of the method by which they are produced and regardless of the number of subunits. It also includes fragments of proteins, peptides and oligopeptides. They may be natural proteins, recombinant proteins or fusion proteins. Proteins of the present invention are preferably composed of at least 5 amino acids, preferably at least 10 amino acids, preferably at least 20 amino acids. It is understood that proteins useful in the present invention are usually isolated proteins, for example when derived from natural products such as plants.

[0059] The active protein of the present invention is preferably an active protein intended for the pharmaceutical, veterinary, cosmetic, food or nutraceutical fields.It is preferably selected from pharmaceutical proteins, such as enzymes, cytokines, hormones, growth factors, plasma factors, vaccines, antibodies.Preferably, the pharmaceutical protein useful in the present invention is insulin or a pharmacologic active derivative thereof, preferably insulin.

[0060] Therefore, in a further aspect, the present invention relates to a composition intended for oral administration of a biologically active substance, said composition comprising cross-linked maltodextrin and an effective amount of said biologically active substance for use as a medicament.

[0061] When the biologically active substance is preferably insulin and / or a pharma- ceutical active derivative thereof, the present invention also refers to a composition intended for oral administration of insulin and / or a pharmaceutical derivative thereof, comprising cross-linked maltodextrin and an effective amount of insulin and / or a pharmaceutical derivative thereof, for use in the prevention or treatment of diabetes, preferably type 1 diabetes and / or gestational diabetes.

[0062] The uses according to the invention encompass pharmaceutical or non-pharmaceutical uses, which essentially depend on the biological activity involved and / or the condition to be treated and / or prevented by said biological activity.

[0063] The present invention therefore relates to a composition intended for the oral administration of a biologically active substance, comprising cross-linked maltodextrin and an effective amount of said biologically active substance.

[0064] Preferably, the cross-linked maltodextrin is similar to that previously described in the preferred embodiment.

[0065] Preferably, the biologically active substance is as previously described in the preferred embodiments, it is preferably insulin and / or its pharma- ceutical active derivatives.

[0066] Preferably, the composition of the present invention is a medicament for the treatment or prevention of diabetes, preferably type 1 diabetes and / or gestational diabetes. Even more preferably, it is for the treatment of diabetes, preferably type 1 diabetes and / or gestational diabetes.

[0067] In the compositions of the present invention, the biologically active agent is advantageously loaded into the cross-linked maltodextrin.

[0068] To carry out the loading, the crosslinked maltodextrin of the present invention can be used in liquid, solid or semi-solid state. For example, the crosslinked maltodextrin can be mixed with a small amount of water to obtain a gel. This gel is then mixed with the biologically active substance to be loaded, either in powder form or dissolved in a suitable solvent, by kneading and / or mixing. Alternatively, the loaded crosslinked maltodextrin can be easily obtained by adding a selected amount of the crosslinked maltodextrin together with an excess of the guest biologically active substance dissolved in a suitable solvent after stirring overnight at room temperature. The loading occurs and is recovered by simply filtering under vacuum.

[0069] Therefore, the present invention further relates to a method for the preparation of a composition according to the invention, comprising the step of loading the crosslinked maltodextrin useful in the present invention with an effective amount of a biologically active substance.

[0070] The present invention also relates to a method for treating an individual comprising oral administration of a composition according to the present invention.

[0071] Preferably, the method is for the treatment or prevention of diabetes, more preferably type 1 diabetes and / or gestational diabetes. Even more preferably, the method is for the treatment of diabetes, preferably type 1 diabetes and / or gestational diabetes.

[0072] Preferably, the method is for treating an individual suffering from diabetes, preferably type 1 diabetes and / or gestational diabetes. EXAMPLES

[0073] The invention will be better understood in light of the following examples.

[0074] example A. Delivery system synthesis 1. Delivery system according to the invention (MDX-PYR): maltodextrin cross-linked with pyromellitic dianhydride (PMDA) / anhydroglucose units molar ratio of 0.57.

[0075] Into a round bottom flask set on a water bath and magnetic stirrer, 40 mL of DMSO containing 9.78 g of dehydrated KLEPTOSE® Linecaps (smooth pea starch derived from maltodextrin with an amylose content of 35-45%, DE of 17, and weight average molecular weight of 12000 Da) was introduced and stirred until a colorless homogeneous solution was obtained. 10 mL of triethylamine was added to the solution, followed by 7.52 g of pyromellitic dianhydride. A dark amorphous solid-like gel formed and was left to solidify overnight. Using a mortar and pestle, the crosslinked maltodextrin was ground, washed several times with distilled water, then acetone in a Büchner, dried, and then extracted with acetone in a Soxhlet at 60-70 °C for 48 h. Finally, the clean powder was dried, ground, and collected.

[0076] The suspension of MDX-PYR particles thus obtained was prepared using the top-down method. MDX-PYR was suspended in saline (NaCl 0.9% w / v) at a concentration of 10 mg / ml under distillation at room temperature. The suspension was then dispersed for 10 min using a high-shear homogenizer (Ultraturrax®, IKA, Konigswinter, Germany). The suspension was then subjected to high-pressure homogenization for 90 min at 500 bar back pressure (EmulsiFlex C5, Avastin, USA). Finally, the MDX-PYR suspension was clarified by dialysis to remove potential synthetic residues (Spectrapol, cellulose membrane, cut-off 12000 Da) and finally stored at -4 °C.

[0077] 2. Comparative delivery system (BCD-PYR): Cyclodextrin cross-linked with pyromellitic dianhydride (PMDA) / anhydroglucose units molar ratio of 0.57. Comparative crosslinked β-cyclodextrin BCD-PYR was prepared according to Example 1 of Italian patent application ITBO20120710.

[0078] B. Characterization of the delivery system The delivery systems obtained according to section A above were characterized for the following characteristics: mean diameter, polydispersity index, zeta potential, pH value.

[0079] The pH was assessed at room temperature for suspensions containing 10 mg / mL of the delivery system using a pH meter (Orion model 420A).

[0080] For the determination of mean diameter and polydispersity index and zeta potential, a suspension containing 10 mg / mL of the delivery system was first diluted with filtered (0.22 μm) distilled water using a dilution factor of 1 / 30 volume.

[0081] The mean diameter and polydispersity index were determined by laser light scanning (90plus Instrument, Brookhaven, NY, USA) on a dilute suspension of the delivery system. The analysis was carried out at a scattering angle of 90° at a temperature of 25° C., with a refractive index of 1.330 and a viscosity of 0.890 cPs.

[0082] Zeta potential was measured on dilute suspensions of the delivery system by electrophoretic mobility using the same instrument. The analysis was performed at a scattering angle of 90° C. and a temperature of 25° C. For the zeta potential measurements, a sample of the dilute delivery system formulation was placed in an electrophoretic cell and an electric field of 15 V / cm was applied.

[0083] The results are shown in Table 1.

[0084] [Table 1]

[0085] C. Insulin loading 1. Insulin Loading Protocol Insulin from bovine pancreas powder was used to prepare a 2 mg / mL solution in distilled water adjusted to pH 2.3 with phosphoric acid. The insulin solution was added to the preformed aqueous nanosuspension in a mass ratio of 1:5 insulin solution:nanosuspension. The mixture was stirred at room temperature for 30 min, incorporated, centrifuged, and then the supernatant separated from the precipitate was collected and lyophilized for future use.

[0086] 2. evaluation A. Loading capacity The loading capacity was determined from the lyophilized insulin-loaded samples. Briefly, 2-3 mg of insulin-loaded lyophilized delivery system weight was dispersed in 5 mL of distilled water. Sonication (15 min, 100 W) and centrifugation treatment were performed to allow the release of insulin from the delivery system. The supernatant was then analyzed for quantitative measurement of insulin.

[0087] Quantification of insulin was performed by high performance liquid chromatography analysis with an HPLC pump (Perkin Elmer 250B, Waltham, MA) equipped with a spectrophotometer detector (Flexar UV / Vis LC, Perkin Elmer, Waltham, MA). An analytical column C18 (250 mm × 4.6 mm, ODS Ultrasphere 5 μm; Beckman Instruments, USA) was used. The mobile phase consisted of a mixture of 0.1 M sodium sulfate and acetonitrile (72:28 v / v) in distilled water, filtered through a 0.45 μm nylon membrane and ultrasonically degassed before use. UV detection was fixed at 214 nm and the flow rate was set at 1 ml / min. Insulin concentrations were calculated using the external standard method from a standard calibration curve. For this, 1 mg of insulin was weighed, placed in a 10 mL volumetric flask and dissolved in distilled water adjusted to pH 2.3 with phosphoric acid to obtain a mother solution. This solution was then diluted with the mobile phase to prepare a series of standard solutions, which were then injected into the HPLC system. A linear calibration curve was obtained over the concentration range of 0.5-25 μg / mL, which was plotted linearly with a regression coefficient of 0.999.

[0088] The loading capacity of the delivery system was calculated as follows: [weight of insulin / weight of lyophilized delivery system] x 100.

[0089] The results are shown in Table 2.

[0090] [Table 2]

[0091] b. In vitro drug release kinetics In vitro drug release experiments were performed in a multicompartment rotating disk (a diffusion cell system containing a donor chamber separated by a membrane from the donor compartment), consisting of several donor cells on one side separated from recipient cells on the other side by a cellulose membrane (Spectrapore, cut-off 12000-14000 Da). The insulin-loaded delivery system was placed in the donor cells (1 mL). The recipient cells were filled separately with phosphate-buffered saline PBS solutions (pH 1.2 and pH 6.8). The in vitro release study was performed for 6 h, and the recipient phase was removed at regular intervals and replaced with the same amount of fresh PBS solution. The sampling times investigated were 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 5, and 6 h. The insulin concentration in the collected samples was later detected by HPLC.

[0092] The results are shown in Figures 1 and 2.

[0093] Like the prior art delivery systems, the inventive delivery system MDX-PYR prevents insulin release at respiratory pH (Figure 1) while allowing insulin release at intestinal pH (Figure 2). That is, the inventive delivery system MDX-PYR prevents insulin release at pH levels that would be absorbing due to the high acidity of the urinary tract, and allows insulin release in the intestine where insulin absorption is desired.

[0094] However, in contrast to the prior art delivery system BCD-PYR, the delivery system MDX-PYR of the present invention advantageously allows for a slower release of insulin, i.e., insulin is potentially bioavailable for a longer period of time. This also means that the delivery system of the present invention is less likely to cause a brutal increase in blood insulin after ingestion, i.e., the delivery system of the present invention is potentially safer, since a brutal increase in blood insulin can lead to harmful hypoglycemia.

[0095] This great potential of the delivery system of the present invention was investigated and confirmed in the following in vivo experiments.

[0096] c. In vivo oral administration of insulin-loaded delivery system The delivery system formulation was administered to mice by oral gavage in the stomach. The insulin concentration was 6 U / ml (210 mg / ml insulin), so the dose was 30 U / kg and blood samples were collected at 0, 30, 60, 120, 180 and 240 min. Insulin was later extracted from plasma samples using the following protocol: 100 μl of plasma, 100 μl of PBS (pH 7.4), 50 μl of acetonitrile, 20 μl of ethylparaben, 3 ml of dichloromethane / n-hexane (1:1 v / v) were added, respectively. The mixture was vortexed for 2 min and then centrifuged at 5000 rpm for 10 min. The supernatant was transferred to a test tube, evaporated and the organic phase was placed under nitrogen flux. Afterwards, 300 μl of HCl 0,05N was added for 2 min on a vortex. The organic phase was completely evaporated under nitrogen flux and then centrifuged at 15000 rpm for 10 min. The clear supernatant was injected into the HPLC after appropriate dilution.

[0097] The results are shown in Figure 3.

[0098] After oral administration of insulin-loaded cross-linked maltodextrin MDX-PYR of the present invention, blood insulin increases to 4 ppm / 50 μL of blood pressure, then slowly decreases over time. In contrast, when using the prior art cross-linked β-cyclodextrin BCD-PYR, blood insulin shows a brutal increase in blood plasma to 8 ppm / 50 μL, followed by a rapid decrease. Four hours after oral ingestion, blood insulin is two times higher using the delivery system of the present invention.

[0099] These experiments therefore confirmed the advantageous pharmacokinetic behavior provided by the delivery system of the present invention. The results confirmed (i) greater safety (lower blood insulin peak after ingestion) and (ii) longer-lasting effects (higher blood insulin even after 4 hours) associated with the use of the delivery system of the present invention.

Claims

1. 1. A composition intended for oral administration of a biologically active agent comprising cross-linked maltodextrin and an effective amount of a biologically active agent, the composition comprising: the biologically active substance is insulin; The crosslinked maltodextrin is obtained by reacting maltodextrin with a bifunctional crosslinking compound, and the molar ratio between the crosslinking compound for the crosslinking reaction and the anhydroglucose units of the maltodextrin is selected in the range of 0.1 to 10.0; The composition, wherein the maltodextrin is derived from starch having a weight average molecular weight in the range of 5,000-15,000 Da and containing 25-50% amylose, expressed by dry weight, based on the total dry weight of the starch.

2. The composition described in claim 1, wherein the maltodextrin has a dextrose equivalent in the range of 5 to 20.

3. The composition of claim 1 or claim 2, wherein the bifunctional crosslinking compound is selected from a dianhydride.

4. The composition of claim 3, wherein the dianhydride is pyromellitic dianhydride.

5. A composition according to any one of claims 1 to 4 for use as a medicament.

6. 6. The composition of claim 5 for use in the treatment or prevention of diabetes.

7. The composition for use according to claim 6, wherein the diabetes is type 1 diabetes and / or gestational diabetes.

8. A method for preparing a composition according to any one of claims 1 to 4 or a composition for use according to any one of claims 5 to 7, comprising the step of loading said crosslinked maltodextrin with an effective amount of said biologically active substance.

9. A long-term sustained release oral formulation comprising the composition according to any one of claims 1 to 4 and suitable excipients.

10. 10. The sustained release oral formulation of claim 9, wherein the formulation is for use as a pharmaceutical or veterinary product.

11. 10. The sustained release oral formulation of claim 9, wherein the formulation is a product selected from the group consisting of cosmetics, food and dietary supplements.

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