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
  • A61K9/1641—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
  • A61K9/1647—Polyesters, e.g. poly(lactide-co-glycolide)
• • 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
  • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
  • A61K9/0065—Forms with gastric retention, e.g. floating on gastric juice, adhering to gastric mucosa, expanding to prevent passage through the pylorus

Definitions

• the present invention relates to a diagnostic marker for the diagnosis of aspiration and gastroesophageal reflux in children and adults, to the manufacture of such markers and the use thereof.
• Aspiration into the airways is one of the major causes of lung disease in infants and young children. Unfortunately the diagnosis of aspiration is often delayed due to unawareness and to the low sensitivity and specificity of existing diagnostic tests.
• Aspiration of food or gastric material into the tracheobronchial tree can result in a variety of disease states such as laryngeal and tracheo-bronchial inflammation leading to stridor, cough, wheezing, recurrent pneumonia, bronchiectasis, pulmonary fibrosis and even suffocation and death due to massive aspiration (Mendelson CL. The aspiration of stomach contents into the lungs during obstetric anesthesia. Am J Obstet Gynecol 1946;52:191-205; Bauer ML, Figueroa-Colon , Georgeson K, Young DW. Chronic pulmonary aspiration in children. South Med J 1993; 86:789-795; Moran TJ. Experimental aspiration pneumonia. Arch Pathol 1955;60:122-129)
• the common conditions that predispose to aspiration in infants and young children include prematurity due to developmental delay . of the coordination between swallowing and breathing (Harding R, Johnson P, McCleala ⁇ d ME. Liquid sensitive laryngeal receptors in the developing ship cat and monkey. J Physiol Lond 1978;277:409-422), abnormal communication between airways and esophagus such as laryngo-tracheal cleft or tracheo-esophageal fistula, massive gastro-esophageal reflux, and neurological abnormalities such as cerebral palsy, vocal cord paralysis and familial dysautonomia.
• Elidemir and his colleagues (Elidemir O, Fan LL, Colasurdo N. A novel diagnostic method for pulmonary aspiration in a murine model. Immunocytochemical staining of milk proteins in alveolar macrophages. Am J Respir Crit Care Med 2000;161 :622-626) have recently shown that aspiration of milk in mice could be diagnosed by specific immunocytochemical staining of milk proteins in bronchoalveolar macrophages. Although this method seems to be specific for the detection of milk, aspiration of other food substances cannot be detected by this method. Furthermore they stained milk proteins ( ⁇ -lactalbumin and ⁇ -lactoglobulin) in alveolar macrophages by an immunocytochemical method.
• the marker of the present invention is one that is safe and bio-degradable, can be given orally with food, can easily be "swallowed” by alveolar macrophages and can be identified in bronchoalveolar lavage (BAL) alveolar macrophages for a substantial period of time.
• BAL bronchoalveolar lavage
• charcoal particles were not suitable and would not be acceptable as a diagnostic tool for humans due to the danger inherent in introducing charcoal particles into the lungs.
• a diagnostic composition for detecting both aspiration and gastroesophageal reflux comprising bio-degradable microspheres having a diameter of about 0.1 - 10 microns.
• said microspheres have a diameter of about 1 - 4 microns.
• diagnostic composition for detecting both aspiration and gastroesophageal reflux comprising bio-degradable polymeric microspheres having a diameter of about 0.1 - 10 microns.
• Polymers which undergo degradation under these conditions are, among others, polyesters, polyphosphate esters, polyphosphazenes, polyorthoesters, polyanhydrides, polycarbonates, polyamides and proteins (polypeptides).
• Typical polyesters are e.g. homopolymers or different copolymers of lactic acid (or lactide), glycolic acid (or glycolide), mandelic acid, caprolactone, ⁇ - hydroxy acids (e.g. hydroxy butyric acid).
• Polymers containing optically active monomers may consist of only d- or I- monomers, or any combination of the two. By varying the chemical composition and/or the molecular weight of the polymer, it is possible to control its degradation rate.
• a food product containing a diagnostic composition for detecting both aspiration and gastroesophageal reflux said composition comprising bio-degradable microspheres having a diameter of about 0.1 - 10 microns.
• microspheres of polylactic acid are especially preferred for use in the present invention.
• d,l- PLA polylactic acid - Boehringer
• acetone Ferutarom
• 100 ml of distilled water were added dropwise, over a period of 60 minutes, under mild magnetic stirring.
• Spherical particles of PLA 1-2 microns in diameter, are formed at an early stage in the process by precipitation.
• the acetone and part of the distilled water were then evaporated by using a rotary evaporator (Buchi), at 40°C.
• a surfactant Pluronic F-68
• the method is tolerant to relatively large changes in the amount of both the acetone and distilled water.
• microspheres are easily visible by light microscopy, both with and without a polarizer, the possibility of coloring the microspheres was explored.
• the colorants which are added to the acetone solution, should dissolve in acetone and be relatively hydrophobic, in order to be included inside the particles during the precipitation process.
• another indicator such as a fluorescent material may be added.
• microspheres were efficiently suspended (10mg/ml) within sterile normal saline by sonification as shown by light microscopy.
• ketamine HCI 50 mg/ml, 10 ml
• dehydrobenzperidol 2.5 mg/ml, 2 ml
• Tracheal instillation Direct intubation was performed with a blunted metal needle. A few ventilations with a small ambu bag were given to ascertain intratracheal position of the needle by confirming chest movement. Normal saline or microsphere suspension (0.1 ml) was slowly instilled into the trachea, followed by a few ventilations with the ambu bag to disperse the fluid and to prevent apnea.
• Bronchoalveolar lavage On the planned BAL day, under anesthesia, exsanguination was performed by transection of the abdominal aorta. The trachea was exposed and a blunted naedle covered with a polyethylene cannula was inserted into the trachea. Three aliquots of 5 ml sterile 0.9% saline were injected and withdrawn with a total recovery of 85.3 + 0.8 %. The fluid was examined for total cell counts and slides for differential counts were prepared on a Shandon Cytospin 3 (Cheshire, England) using approximately 100 ⁇ L of BAL fluid.
• Microsphere index The first 100 consecutive intact macrophages viewed were evaluated and the total number of white microspheres within the 100 macrophages was defined as the microsphere index on each BAL day.
• Histology Lungs from all animals were preserved in formaldehyde for further evaluation and four slides from both lungs were prepared. Other internal organs (kidneys, liver, spleen, adrenals, pancreas, heart and brain) from 8 hamsters after 90 days of tracheal instillation (5 following white microsphere instillation and 3 following, saline instillation) were preserved in formaldehyde for further evaluation Animals instilled black microspheres had all extrapulmonary organs, including perihilar lymph glands, preserved in formaldehyde for histological evaluation.. Two slides from each extrapulmonary organ were prepared. Hematoxylin-eosin was used for staining. Slides were thoroughly evaluated by two senior pathologists (V.D and Y.S.).
• the white microspheres of poylactic acid are clearly identified within the alveolar macrophages and therefore can be used as a specific sensitive and stable marker for the diagnosis of aspiration.
• non-solvents and surfactants are suitable in the preparation process of the microspheres for use in the present invention.
• proteins which may be used for this purpose are human albumin, hemoglobin, gelatin and soybean proteins.
• Example 2 Precipitation with a non-solvent:
• Example 1 The procedure of Example 1 as described for PLA is repeated, except that the solvent is either water or an aqueous solution and the non-solvent is a water- soluble volatile solvent, such as acetone.
• the solvent is either water or an aqueous solution and the non-solvent is a water- soluble volatile solvent, such as acetone.
• the crosslinking agent can be any diacylchloride, or any di- or poly functional molecule able to react with the proteins chemically active groups. Using this method, either microspheres or microcapsules may be obtained. The rigidity of the microspheres may be controlled by changing the nature and concentration of the crosskinking agent, as well as the nature of the organic phase and rinsing solvents.
• the protein is either precipitated as in Example 2, or emulsified as in Example 3., after which, the suspension is heated sufficiently to induce denaturation of the protein, thus stabilizing the particles.

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• Health & Medical Sciences (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Epidemiology (AREA)
• Medicinal Chemistry (AREA)
• Pharmacology & Pharmacy (AREA)
• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Animal Behavior & Ethology (AREA)
• General Health & Medical Sciences (AREA)
• Public Health (AREA)
• Veterinary Medicine (AREA)
• Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
• Medicinal Preparation (AREA)
• Coloring Foods And Improving Nutritive Qualities (AREA)

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• 2002
  • 2002-07-25 IL IL15090602A patent/IL150906A0/xx not_active IP Right Cessation
• 2003
  • 2003-07-03 JP JP2004524029A patent/JP2005537283A/ja active Pending
  • 2003-07-03 US US10/522,200 patent/US8231860B2/en not_active Expired - Fee Related
  • 2003-07-03 EP EP03771237A patent/EP1575620B1/en not_active Expired - Lifetime
  • 2003-07-03 WO PCT/IL2003/000552 patent/WO2004011035A2/en not_active Ceased
  • 2003-07-03 AU AU2003242957A patent/AU2003242957A1/en not_active Abandoned

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