WO2016174573A1 - Process for producing polysaccharide microparticles for alveolar macrophage targeting, microparticles obtained therein and use thereof - Google Patents
Process for producing polysaccharide microparticles for alveolar macrophage targeting, microparticles obtained therein and use thereof Download PDFInfo
- Publication number
- WO2016174573A1 WO2016174573A1 PCT/IB2016/052353 IB2016052353W WO2016174573A1 WO 2016174573 A1 WO2016174573 A1 WO 2016174573A1 IB 2016052353 W IB2016052353 W IB 2016052353W WO 2016174573 A1 WO2016174573 A1 WO 2016174573A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- microparticles
- polysaccharide
- spray
- μπι
- drying
- Prior art date
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/438—The ring being spiro-condensed with carbocyclic or heterocyclic ring systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4409—Non condensed pyridines; Hydrogenated derivatives thereof only substituted in position 4, e.g. isoniazid, iproniazid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/007—Pulmonary tract; Aromatherapy
- A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
Definitions
- the present invention relates to microencapsulation techniques of drugs, useful in the production of microparticles for the treatment of tuberculosis, using polysaccharide microparticles obtained by single step spray-drying.
- Alveolar macrophages are resident cells of the lung, particularly of the alveolar zone, being directly involved in the progress of some diseases, such as tuberculosis.
- macrophages host the bacteria responsible for the disease ⁇ Mycobacterium tuberculosis) and are therefore a privileged therapeutic target.
- any drug intended at macrophage targeting is required to reach this zone of the respiratory tract.
- Inhalation of drugs aimed at reaching the alveolar zone requires the design of specific carriers that are endowed with the adequate aerodynamic characteristics to enable the desired distribution within the lung. To permit so, an aerodynamic diameter of 0.5 - 3 ⁇ is usually the referred range.
- Polymeric microparticles are the carriers proposed more frequently for this objective of drug inhalation, because their aerodynamic properties can be tailored to fit specific requirements.
- Spray-drying is the most frequently used method to obtain microparticles.
- This is a microencapsulation technique known to produce microparticles with tunable properties depending on the optimisation of the parameters involved in the proper spray-drying procedure, thus fitting the need of permitting tuning the microparticles to exhibit specific characteristics.
- Macrophages exhibit several surface receptors that might be used to mediate targeting to these cells. Actually, these receptors are those involved in the recognition of pathogenic agents.
- the C-type lectin receptors and the scavenger receptors are the two families reported to be involved in most cases of polysaccharide recognition.
- the first family includes the mannose receptor, which has reported ability to recognise mannose, fucose, iV-acetylglucosamine and sulphated sugars, which are present in many polysaccharide substances.
- the second family has demonstrated affinity for several polysaccharides such as chondroitin sulphate, dextran sulphate and fucoidan, among others. This enhanced ability for recognition of certain units, makes the macrophages highly targetable with materials comprising these residues, namely polysaccharides.
- LAURIENZO P. Marine polysaccharides in pharmaceutical applications: an overview in Marine Drugs, 2010; 8 (9) : 2435-2465, describes that alginate microparticles permitted higher bioavailability and reduction of adverse effects when compared with the administration of antitubercular drugs in the free form.
- KUNDAWALA A. et al Preparation of Microparticles containing rifampicin as dry powder formulation: in vitro studies on aerossol performance in American Journal of Pharmtech Research 2(4), 2012, describe the production, by spray-drying, of a dry powder formulation of rifampicin consisting of chitosan microparticles, for an application in tuberculosis therapy.
- Figure 1 shows the microparticle morphology, observed by scanning electron microscopy (SEM) , wherein CRG denotes carrageenan, FUC denotes fucoidan, KGM denotes Konjac glucomannan, LBG denotes locust bean gum and GG denotes guar gum (partially hydrolysed) .
- SEM scanning electron microscopy
- Figure 2 shows comparative uptake of microparticles composed of different polysaccharides and polyvinyl alcohol (PVA, control), by NR8383 cells (rat alveolar macrophages), upon 2 hours exposure to 50 - 60 g/cm 2 of fluorescently-labelled microparticles (mean ⁇ SEM, n > 3), wherein CRG denotes carrageenan, FUC denotes fucoidan, denotes LBG denotes locust bean gum and GG denotes guar gum (partially hydrolysed) .
- CRG denotes carrageenan
- FUC denotes fucoidan
- LBG denotes locust bean gum
- GG denotes guar gum (partially hydrolysed) .
- the invention refers to microparticles produced by a single-step spray-drying process, which are based on selected polysaccharides regarding the objective of alveolar macrophage targeting.
- the parameters of the spray- drying process are optimised to permit the production of microparticles that evidence aerodynamic properties suitable to reach the alveolar zone, where alveolar macrophages reside.
- the microparticles are aimed at providing an inhalable strategy for tuberculosis therapy, thus requiring macrophage targeting.
- unloaded microparticles were produced and used.
- the present invention uses natural materials and, namely polysaccharides, which are absolutely advantageous, because if polysaccharides composed of the mentioned recognisable residues are used, it becomes possible producing targeted drug carriers in a single step, without the need to include targeting ligands at a posterior phase. Additionally, polysaccharides are obtained at a low cost, have high probability of evidencing biocompatibility and are very flexible structures.
- the strategy of designing polysaccharide carriers for inhalable tuberculosis therapy encompasses the single-step production of a carrier that naturally targets the macrophages.
- polysaccharides composing the microparticles were selected for their chemical structure, as this characteristic is the one specifically mediating the active targeting of macrophages.
- Selected polysaccharides include locust bean gum, guar gum, fucoidan, glucomannan, carrageenan, xanthan gum, glucan, dextran sulfate, chondroitin sulfate. All these polysaccharides bear in their structure, as natural components, chemical groups (namely sulphate) or residues (i.e.
- Locust bean gum is a galactomannan, having mannose and galactose residues in an average molar ratio of 4/1.
- Guar gum is a galactomannan, having mannose and galactose residues in an average molar ratio of 2/1.
- Fucoidan has fucose units in its structure and also sulfate groups.
- Glucomannan has glucose and mannose units in its structure, in a molar ratio of 1/1.6.
- the first step consists on the solubilisation of the polysaccharides.
- Each polysaccharide was used individually and the concentration of polymer varied within 1.5 and 2% (w/v) .
- the lower concentration mentioned (1.5%) was used when 2% was too viscous to enable spray-drying.
- Locust bean gum required grinding before solubilisation.
- Locust bean gum required heating at 85 °C for 30 minutes to solubilise.
- Carrageenan required heating at 50 °C for 30 minutes to solubilise.
- the spray-dryer was used in open mode configuration and compressed air was the applied gas.
- the spray-drying conditions were variable, depending on the material being used.
- the conditions were: inlet temperature between 140 °C and 180 °C, aspirator between 80% and 90%, solutions feed rate between 0.7 and 2 mL/min.
- microparticles either evidenced a spherical shape or a convoluted shape ( Figure 1) .
- Figure 1 microparticles of locust bean gum, guar gum, fucoidan, carrageenan and glucomannan were produced to demonstrate the concept.
- the characteristics of microparticles are summarised in Table 1. Feret diameters were determined to be between 1.4 and 1.8 ⁇ , while real densities ranged between 1.0 and 1.7 g/cm 3 , which resulted in theoretical aerodynamic diameters varying within 1.7 ⁇ and 2.3 ⁇ . Overall, the characteristics indicate the theoretical suitability for deep lung inhalation.
- microparticles were tested for their ability to encapsulate two different first-line antitubercular drugs in association, isoniazid and rifabutin, as required in tuberculosis therapy (single therapy is not recommended) .
- the encapsulation efficiencies varied between 74 and 89% for isoniazid, and 57 and 92% for rifabutin .
- the first object of the invention is a process for producing polysaccharide microparticles for alveolar macrophage targeting, using single step spray-drying, comprising : a) dissolving the polysaccharide in order to obtain a concentration of polymer between 1.5 and 2% (w/v) ;
- the polysaccharide is selected from the group consisting of locust bean gum, fucoidan, guar gum, carrageenan, glucomannan, xanthan gum, glucan, dextran sulfate and chondroitin sulfate.
- Particularly preferred polysaccharides are locust bean gum, fucoidan, guar gum and carrageenan.
- the spray-dryer is normally used in open mode configuration and the the gas applied by the spray-dryer is compressed air.
- the second object of the invention consists of polysaccharide microparticles for alveolar macrophage targeting, obtained by the process described above, having Feret diameters between 1.0 and 2.0 ⁇ , real densities between 1.0 and 1.7 g/cm 3 and theoretical aerodynamic diameters between 1.7 ⁇ and 2.3 ⁇ .
- the polysaccharide microparticles have Feret diameters between 1.4 and 1.8 ⁇ .
- the third object of the invention consists of such polysaccharide microparticles for use in the treatment of tuberculosis, as carriers for drugs used for treating such disease.
- One preferred drug combination is an association of isoniazid and rifabutin.
- Locust bean gum (LBG) microparticles 1 g of LBG is grinded in a mortar for 10 min, after which 5 mL HC1 0.1 M are added, while grinding continues until complete mixture of LBG and HC1. Purified water previously heated to 85 °C is then added, up to a final volume of 50 mL (LBG concentration is 2%, w/v) . The dispersion is maintained under magnetic stirring for 30 min and subsequently placed on a water bath at 85 °C under slow stirring for additional 30 min. At the end, the dispersion is kept under stirring at room temperature overnight, until the moment of spray- drying. Spray-drying operating parameters are: inlet temperature: 160 °C; aspirator: 85%; feed rate: 0.8 mL/min; and spray flow rate: 473 L/h. Microparticle production yield is 70%.
- GG microparticles 1 g of GG (partially hydrolysed) is dissolved in 50 mL of purified water under stirring for 20 min (GG concentration is 2%, w/v) . After dissolution, mannitol is added and dissolved in the GG solution to a final concentration of 0.5% (m/v) , in order to improve the final aerodynamic characteristics of the microparticles. The final solution is maintained under stirring for 20 min before spray-drying. Spray-drying operating parameters are: inlet temperature: 160 ⁇ 2 °C; aspirator: 80%; feed rate: 1.0 mL/min; and spray flow rate: 473 L/h. Microparticle production yield is 73%.
- KGM (partially hydrolysed) is dissolved in 50 mL of purified water, at 70 °C, under stirring for 45 min (KGM concentration is 2%, w/v) .
- Spray-drying operating parameters are: inlet temperature: 170 ⁇ 2 °C; aspirator: 90%; feed rate: 0.7 mL/min; and spray flow rate: 473 L/h. Microparticle production yield is 69%.
- THP-1 cells might be differentiated (using phorbolmyristate acetate, PMA) to exhibit macrophage phenotype and develop characteristics of alveolar macrophages.
- NR8383 cells are rat alveolar macrophages.
- Alveolar macrophages have scavenger receptors on the surface, including the mannose receptor (CD206) , reported as capable of recognising mannose, fucose units and sulphated sugars.
- CD206 mannose receptor
- the differentiation of THP-1 cells by ⁇ is reported to not induce these receptors, while they are described to be present in NR8383 cells. While all the selected polysaccharides display structural units capable of undergoing specific recognition by macrophages, polyvinyl alcohol (PVA) is devoid of such moieties and was used as control.
- PVA polyvinyl alcohol
- each polysaccharide was previously labelled with a fluorescent marker before the spray-drying process.
- Microparticles composing each formulation were insufflated over cell layers in 6-well plates and incubated for 2 hours. After that period, the cells were recovered and analysed by flow cytometry, to examine the compositions favouring macrophage capture .
- Locust bean gum, fucoidan, guar gum and carrageenan were the polysaccharides undergoing the experimental test of uptake, apart from the control PVA.
- Flow cytometry determines the number of macrophages in the sample and quantifies the amount of macrophages that exhibit fluorescence, which is a consequence of having phagocytosed microparticles that are fluorescently labelled.
- a control test was performed using non- fluorescent microparticles. In that case, no macrophages are detected as emitting a fluorescent signal or the detected signal is not significant (maximum obtained was 1%) .
- NR8383 cells give the most meaningful results, as this is the cell line exhibiting the receptor of interest.
- locust bean gum is the polymer showing higher affinity for macrophages, inducing 95% capture, followed by guar gum and carrageenan, which have a similar result (around 70% capture) .
- Curiously the uptake observed for fucoidan microparticles did not overpass 30%, inclusive remaining at a lower level comparing with the control PVA.
- the increase of the dose of contact to 220-250 g/cm 2 resulted in a general increase to values around 100% for all polysaccharides, indicating a dose-dependent effect (data not shown) .
- Locust bean gum, guar gum and the control PVA were also tested in macrophage-differentiated THP-1 cells at the concentration of 50-60 g/cm 2 , which resulted in an increase of capture to values around 90-100% in all cases, possibly as a consequence of the absence of the specific mannose receptor that could differentiate the uptake. This indicates that the presence of such receptor indeed enables a specific targeting .
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Otolaryngology (AREA)
- Pulmonology (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP16729627.6A EP3288537A1 (en) | 2015-04-29 | 2016-04-26 | Process for producing polysaccharide microparticles for alveolar macrophage targeting, microparticles obtained therein and use thereof |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PT108410 | 2015-04-29 | ||
PT10841015 | 2015-04-29 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2016174573A1 true WO2016174573A1 (en) | 2016-11-03 |
Family
ID=56132975
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/IB2016/052353 WO2016174573A1 (en) | 2015-04-29 | 2016-04-26 | Process for producing polysaccharide microparticles for alveolar macrophage targeting, microparticles obtained therein and use thereof |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP3288537A1 (pt) |
PT (1) | PT116887B (pt) |
WO (1) | WO2016174573A1 (pt) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1997035562A1 (en) * | 1996-03-23 | 1997-10-02 | Danbiosyst Uk Limited | Polysaccharide microspheres for the pulmonary delivery of drugs |
WO2010007604A2 (en) * | 2008-07-16 | 2010-01-21 | Royal College Of Surgeons In Ireland | Inhalable microparticles, and methods for the production thereof |
-
2016
- 2016-04-26 WO PCT/IB2016/052353 patent/WO2016174573A1/en unknown
- 2016-04-26 PT PT116887A patent/PT116887B/pt active IP Right Grant
- 2016-04-26 EP EP16729627.6A patent/EP3288537A1/en not_active Withdrawn
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1997035562A1 (en) * | 1996-03-23 | 1997-10-02 | Danbiosyst Uk Limited | Polysaccharide microspheres for the pulmonary delivery of drugs |
WO2010007604A2 (en) * | 2008-07-16 | 2010-01-21 | Royal College Of Surgeons In Ireland | Inhalable microparticles, and methods for the production thereof |
Non-Patent Citations (4)
Title |
---|
ALIASGAR J. KUNDAWALA ET AL: "Influence of Formulation Components on Aerosolization Properties of Isoniazid Loaded Chitosan Microspheres", INTERNATIONAL JOURNAL OF PHARMACEUTICAL SCIENCES AND DRUG RESEARCH, 1 October 2011 (2011-10-01), pages 297 - 302, XP055286371, Retrieved from the Internet <URL:http://www.ijpsdr.com/pdf/vol3-issue4/5.pdf> * |
GRENHA A. ET AL.: "Natural carriers for application in tuberculosis treatment", JOURNAL OF MICROENCAPSULATION: MICRO AND NANO CARRIERS, vol. 30, no. 3, 2013, XP055286024, DOI: doi:10.3109/02652048.2012.726283 |
KUNDAWALA A. ET AL.: "Preparation of Microparticles containing rifampicin as dry powder formulation: in vitro studies on aerossol performance", AMERICAN JOURNAL OF PHARMTECH RESEARCH, vol. 2, no. 4, 2012 |
LAURIENZO P.: "Marine polysaccharides in pharmaceutical applications: an overview", MARINE DRUGS, vol. 8, no. 9, 2010, pages 2435 - 2465 |
Also Published As
Publication number | Publication date |
---|---|
PT116887A (pt) | 2021-03-31 |
EP3288537A1 (en) | 2018-03-07 |
PT116887B (pt) | 2022-03-09 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Algharib et al. | Preparation of chitosan nanoparticles by ionotropic gelation technique: Effects of formulation parameters and in vitro characterization | |
Arya et al. | Electrospraying: a facile technique for synthesis of chitosan‐based micro/nanospheres for drug delivery applications | |
Pourshahab et al. | Preparation and characterization of spray dried inhalable powders containing chitosan nanoparticles for pulmonary delivery of isoniazid | |
Hou et al. | Preparation and characterization of novel drug-inserted-montmorillonite chitosan carriers for ocular drug delivery | |
Masalova et al. | Alginate and chitosan gel nanoparticles for efficient protein entrapment | |
WO2015192603A1 (zh) | 一种不含表面活性剂的水包油乳液及其用途 | |
Yang et al. | Dry powder inhaler formulation of lipid–polymer hybrid nanoparticles via electrostatically-driven nanoparticle assembly onto microscale carrier particles | |
Young et al. | Positively and negatively surface-charged chondroitin sulfate-trimethylchitosan nanoparticles as protein carriers | |
Doan et al. | Preparation of rifampicin-loaded PLGA microspheres for lung delivery as aerosol by premix membrane homogenization | |
Chen et al. | Preparation of methotrexate-loaded, large, highly-porous PLLA microspheres by a high-voltage electrostatic antisolvent process | |
Thien | Electrospun chitosan/PVA nanofibers for drug delivery | |
US20040071780A1 (en) | PACE-A microspheres for delivery of antigens | |
WO2012160088A1 (de) | Biokompatible nano- und mikropartikel beschichtet mit stabilisatoren für die pulmonale applikation | |
Rodriguez et al. | Effect of formulation and process parameters on chitosan microparticles prepared by an emulsion crosslinking technique | |
CN105749296A (zh) | 一种溃疡性结肠炎组织靶向分子及其应用 | |
Jatal et al. | Lung targeted electrosprayed chitosan nanocomposite microparticles boost the cytotoxic activity of magnolol | |
Ghasemishahrestani et al. | Tunable synthesis of gelatin nanoparticles employing sophorolipid and plant extract, a promising drug carrier | |
Carvalho et al. | Development of pullulan‐based carriers for controlled release of hydrophobic ingredients | |
Ahmady et al. | Alginate carrier as a controlled thymol delivery system: Effect of particle size | |
WO2016174573A1 (en) | Process for producing polysaccharide microparticles for alveolar macrophage targeting, microparticles obtained therein and use thereof | |
Lu et al. | Antheraea pernyi silk fibroin nanoparticles for drug delivery | |
Kaur et al. | Formulation and development of colon-targeted mucopenetrating metronidazole nanoparticles | |
Rodrigues et al. | Investigating the Effect of Chitosan’s Degree of Deacetylation on Size of the Nanoparticle | |
Albetawi | Current Research on Spray-Dried Chitosan Nanocomposite Microparticles for Pulmonary Drug Delivery | |
Johari et al. | Comparison Study Between Encapsulation of Acalypha indica Linn Extracts with Chitosan-PCL and Chitosan-OA |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 16729627 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |