WO2023094987A1 - A drug delivery composition for treating myocardial infraction (mi) - Google Patents

A drug delivery composition for treating myocardial infraction (mi) Download PDF

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Publication number
WO2023094987A1
WO2023094987A1 PCT/IB2022/061276 IB2022061276W WO2023094987A1 WO 2023094987 A1 WO2023094987 A1 WO 2023094987A1 IB 2022061276 W IB2022061276 W IB 2022061276W WO 2023094987 A1 WO2023094987 A1 WO 2023094987A1
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Prior art keywords
exemplary
plga
peg
crgd
platelet
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PCT/IB2022/061276
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French (fr)
Inventor
Aliyeh GHAMKHARI
Farhang Abbasi
Hossein AHMADI TTAFTI
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Ghamkhari Aliyeh
Farhang Abbasi
Ahmadi Ttafti Hossein
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Application filed by Ghamkhari Aliyeh, Farhang Abbasi, Ahmadi Ttafti Hossein filed Critical Ghamkhari Aliyeh
Publication of WO2023094987A1 publication Critical patent/WO2023094987A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
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    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
    • A61K38/1825—Fibroblast growth factor [FGF]
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    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0028—Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
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    • A61K47/6901—Conjugates being cells, cell fragments, viruses, ghosts, red blood cells or viral vectors
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    • A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
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Definitions

  • the present disclosure generally relates to a drug delivery composition for healing myocardial infarction (MI), and more particularly, relates to a method of producing and administering a drug delivery composition for healing MI.
  • MI myocardial infarction
  • MI Myocardial infarction
  • HF heart failure
  • bFGF basic fibroblast growth factor
  • Direct injection of bFGF into heart has many drawbacks, i.e., health risks which can limit its clinical application.
  • different drug delivery systems have been introduced such as systematic drug delivery.
  • Systemic drug delivery is a method of administering a drug, a nutrition, etc., so that whole body is affected. Systemic drug delivery has low efficiency because of slow tissue penetration, short in-vivo half-life, and tendency to cause systemic side effects.
  • an exemplary drug delivery composition for healing myocardial infarction (MI).
  • an exemplary drug delivery composition may include a plurality of core-shell structured microbubbles and a polyvinyl alcohol (PVA) solution.
  • each respective core- shell structured microbubble may include a core and a shell surrounding an exemplary core.
  • an exemplary core may include liquid perfluorohexane (PFH).
  • an exemplary shell may include poly(D,L-lactide-co-glycolide)-heparin-polyethylene glycol-cyclic arginine glycine aspartate - platelet (PLGA-HP-PEG-cRGD- platelet), basic fibroblast growth factor (bFGF) attached to heparin of PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:10000 to 1:10 (bFGF: PLGA-HP-PEG-cRGD-platelet), at least one stabilizing agent attached to PLGA-HP- PEG-cRGD- platelet, and at least one fluorescent label attached to bFGF.
  • PLGA-HP-PEG-cRGD- platelet poly(D,L-lactide-co-glycolide)-heparin-polyethylene glycol-cyclic arginine glycine aspartate - platelet
  • bFGF basic fibroblast growth factor
  • an exemplary liquid PFH to an exemplary PLGA-HP-PEG-cRGD-platelet may have a weight ratio in a range of 1:10 to 1:1 (liquid PFH: PLGA-HP-PEG-cRGD-platelet).
  • an exemplary plurality of core-shell structured microbubbles may be dispersed in an exemplary PVA solution.
  • an exemplary at least one stabilizing agent may include at least one of a plurality of gold nanoparticles (AuNPs), polyoxyethylene (20) sorbitan monooleate, and combinations thereof.
  • an exemplary at least one stabilizing agent may include a plurality of AuNPs may be surrounded by chitosan (AuNPs-chitosan).
  • each respective AuNP-chitosan may include an AuNP surrounded with a layer of chitosan with a thickness in a range of 10 nm to 100 nm.
  • each respective AuNP of an exemplary plurality of AuNPs may have a particle size in a range of 25 nm to 75 nm.
  • an exemplary drug delivery composition may include an exemplary plurality of AuNPs- chitosan to an exemplary PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:1000 to 1:10 (Au NPs-chitosan: PLGA-HP-PEG-cRGD-platelet).
  • an exemplary drug delivery composition may include polyoxyethylene (20) sorbitan monooleate to an exemplary PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:10 to 1:1 (polyoxyethylene (20) sorbitan monooleate: PLGA-HP-PEG-cRGD- platelet).
  • an exemplary PVA solution may include an aqueous PVA solution with a concentration of PVA in a range of 0.1 w/w% to 5 w/w%.
  • an exemplary at least one fluorescent label may include at least one of fluorescein isothiocyanate, rhodamine, and combinations thereof.
  • each respective core-shell structured microbubble of an exemplary plurality of core-shell structured microbubbles may have a size in a range of 1 pm to 30 pm.
  • an exemplary method may include forming an emulsion of a plurality of core- shell structured microbubbles in which each respective core- shell structure microbubble may have a fluorescent label, injecting a predetermined amount of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles intravenously to a body of a patient, tracking a fluorescent label of each respective core-shell structure microbubble utilizing a fluorescent detector for a time period of maximum 72 hours, and applying an ultrasonic energy to chest of an exemplary patient with acoustic intensity in a range of 1 mWcm” 2 to 600 mWcm" 2 .
  • injecting an exemplary predetermined amount of an exemplary emulsion of an exemplary plurality of core- shell structured microbubbles intravenously to an exemplary body of an exemplary patient may include injecting an amount of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles in a range of 10 pg/mL to 500 pg/mL intravenously to the body of the patient.
  • applying an exemplary ultrasonic energy an exemplary chest of an exemplary patient may include applying an exemplary ultrasonic energy with an acoustic frequency in a range of 1 MHz to 2 MHz to an exemplary chest of an exemplary patient.
  • applying an exemplary ultrasonic energy to an exemplary chest of an exemplary patient may include applying an exemplary ultrasonic energy to an exemplary chest of an exemplary patient after reaching an exemplary drug delivery composition to an exemplary target place, in which an exemplary target place may include platelet-clogged arteries that supply blood to heart.
  • applying an exemplary ultrasonic energy may include applying an exemplary ultrasonic energy to an exemplary chest of an exemplary patient for a time period in a range of 1 minute to 30 minutes.
  • forming an exemplary emulsion of an exemplary plurality of core- shell structured microbubbles may include forming a shell portion and forming a core portion.
  • forming an exemplary shell portion may include forming poly(D,L-lactide-co-glycolide)-NHS (PLGA-NHS) by dissolving PLGA-COOH, l-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), and NHS in an anhydrous dichloromethane solution, forming PLGA-NH2 by dissolving formed PLGA-NHS and ethylenediamine (EDA) in a dichloromethane solution, forming PLGA-HP by mixing dissolved heparin salt, NHS, and EDC with an exemplary formed PLGA-NH2in N,N- dimethylformamide/formamide (DMF/FA), forming HOOC-PEG-COOH by dissolving
  • injecting an exemplary predetermined amount of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles may include injecting an amount of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles in a range of 10 pg/mL to 2000 pg/mL intravenously to an exemplary body of an exemplary patient.
  • forming PLGA-HP may include mixing dissolved heparin salt, NHS, and EDC with an exemplary formed PLGA-NH2 in DMF/FA at a temperature in a range of 4°C to 25°C for a time period in a range of 100 minutes to 2000 minutes.
  • injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution into an exemplary PVA solution may include injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution with a flow rate in a range of 100 pl/min to 500 pl/min into the PVA solution using an exemplary first syringe pump.
  • injecting an exemplary liquid PFH into an exemplary PVA solution may include injecting an exemplary liquid PFH into an exemplary PVA solution with a flow rate in a range of 1 pl/min to 500 pl/min using an exemplary second syringe pump.
  • forming an exemplary PLGA-HP-PEG-cRGD-platelet may include mixing an exemplary platelet solution with an exemplary PLGA-HP-PEG-cRGD with a volume ratio in a range of 100:1 to 10:1 (PLGA-HP-PEG-cRGD: platelet).
  • forming an exemplary PLGA-HP-PEG-cRGD may include mixing an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA with an exemplary cRGD solution with a weight ratio in a range of 10000:1 to 100:1 (PLGA-HP-PEG-NHS: cRGD).
  • FIG. 1A illustrates a flowchart of a method for healing myocardial infarction (MI), consistent with one or more exemplary embodiments of the present disclosure
  • FIG. IB illustrates a flowchart of a method to produce an exemplary plurality of coreshell structured microbubbles, consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 2A illustrates a chemical reaction for synthesizing PLGA-NHS, consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 2B illustrates a chemical reaction for synthesizing PLGA-NH2, consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 2C illustrates a chemical reaction for synthesizing PLGA-HP, consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 2D illustrates a chemical reaction for synthesizing PLGA-HP-PEG-cRGD, consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 3 illustrates a microfluidic device for producing an exemplary plurality of coreshell structured microbubbles, consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 4 illustrates a spectrum of proton nuclear magnetic resonance (1HNMR) spectroscopy of PLGA-HP-PEG-NHS, consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 5 illustrates an optical microscopy image of microbubbles (MBs) obtained at flow rates of 1 pl/min for PFH and 250 pl/min for an exemplary bFGF loaded PLGA-HP-PEG- cRGD-platelet solution, consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 6A illustrates an image of MBs before applying LIFU, consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 6B illustrates an image of MBs after 20 minutes of applying LIFU with acoustic intensity of 35 mW/cm 2 , consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 6C illustrates an image of MBs after 45 minutes of applying LIFU with acoustic intensity of 35 mW/cm 2 , consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 6D illustrates an image of MBs after 7 minutes of applying LIFU with acoustic intensity of 330 mW/cm 2 , consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 6E illustrates an image of MBs after 12 minutes of applying LIFU with acoustic intensity of 330 mW/cm 2 , consistent with one or more exemplary embodiments of the present disclosure
  • FIG. 7 illustrates fluorescence microscopic images of drug delivery samples for analyzing cellular uptake, consistent with one or more exemplary embodiments of the present disclosure.
  • FIG. 8 illustrates fluorescence microscopic images of an exemplary plurality of fluorescent labeled core-shell structured microbubbles and an exemplary plurality of core-shell structured microbubbles with no label, consistent with one or more exemplary embodiments of the present disclosure.
  • the present disclosure is directed to exemplary embodiments of a drug delivery composition for healing myocardial infarction (MI) and a method to produce and administer an exemplary drug delivery composition.
  • MI is a cardiovascular disease that is caused by a blockage in arteries that supply blood to heart.
  • MI is a condition of heart failure due to insufficient blood (oxygen) supply to heart.
  • an exemplary drug delivery composition may carry a drug to a target place in body.
  • an exemplary target place may include platelet-clogged arteries that supply blood to heart.
  • an exemplary drug delivery composition may be activated by an ultrasonic energy to release an exemplary drug.
  • an exemplary drug may include basic fibroblast growth factor (bFGF).
  • an exemplary ultrasonic energy may include a high-intensity focused ultrasound (HIFU) energy and a low-intensity focused ultrasound (LIFU) energy.
  • HIFU high-intensity focused ultrasound
  • LIFU low-intensity focused ultrasound
  • an exemplary LIFU energy may be used due to potential of an exemplary LIFU energy to focus on a specific site, minimize adverse side effects, and prevent damage to surrounding tissues of an exemplary target place.
  • exemplary adverse side effects may include producing undesired heating by HIFU energy.
  • an exemplary ultrasonic energy may be produced using an ultrasonic device applying an exemplary ultrasonic energy with acoustic frequency in a range of 1 MHz to 2 MHz.
  • an exemplary ultrasonic energy may be produced using an ultrasonic device applying an exemplary ultrasonic energy with acoustic intensity in a range of 1 mWcm" 2 to 600 mWcm" 2 .
  • an exemplary drug delivery composition may include a plurality of core-shell structured microbubbles.
  • an exemplary plurality of core-shell structured microbubbles may include a core portion and a shell portion.
  • an exemplary core portion may include liquid perfluorohexane (PFH).
  • PFH liquid perfluorohexane
  • liquid PFH may have a boiling point in a range of 58°C to 60°C.
  • an exemplary liquid PFH may be vaporized via an exemplary ultrasonic energy.
  • an exemplary ultrasonic energy may be produced utilizing an ultrasonic device.
  • an exemplary shell portion of each respective core-shell structured microbubble may include poly(D,L- lactide-co-glycolide)-heparin-polyethylene glycol-cyclic arginine glycine aspartate-platelet (PLGA-HP-PEG-cRGD-platelet), bFGF, at least a fluorescent label, and at least a stabilizing agent.
  • bFGF may be attached to an exemplary heparin of PLGA- HP-PEG-cRGD-platelet.
  • heparin may enhance bFGF loading on PLGA-HP-PEG-cRGD-platelet by interacting with bFGF.
  • heparin may have a negative electrical charge.
  • bFGF may have a positive electrical charge.
  • an ionic interaction may be formed between an exemplary negative electric charge of heparin and an exemplary positive electrical charge of bFGF.
  • RGD is a peptide that may be attached to GP Ilb/IIIa receptors.
  • GP Ilb/IIIa receptors may be formed on surface of activated platelets in blood vessels.
  • cyclic RGD (cRGD) may have greater affinity than linear RGD to GP Ilb/IIIa receptors.
  • an exemplary greater affinity of cRGD in comparison to linear RGD to GP Ilb/IIIa receptors may result in more stability of GP Ilb/IIIa receptor and cRGD.
  • platelet in PLGA-HP-PEG-cRGD-platelet may also increase accumulation of an exemplary plurality pf core-shell structured microbubbles at an exemplary target place.
  • platelet may have physical interaction with activated platelet in an exemplary target place.
  • PEG and PLGA may be biocompatible polymers for carrying bFGF, platelet, and cRGD.
  • dosage of bFGF may be monitored by controlling an amount of bFGF loaded on PLGA-HP- PEG-cRGD-platelet.
  • an exemplary plurality of core- shell structured microbubbles may be dispersed in a polyvinyl alcohol (PVA) aqueous solution to form an emulsion.
  • PVA polyvinyl alcohol
  • an exemplary at least a stabilizing agent may be added to an exemplary shell portion for enhancing stability of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution.
  • an exemplary at least a stabilizing agent may include one of a plurality of gold nanoparticles (AuNPs), polyoxyethylene (20) sorbitan monooleate, and combinations thereof.
  • PVA may also enhance stability of an exemplary plurality of core- shell structured microbubbles in water.
  • an exemplary at least a stabilizing agent may include a plurality of AuNPs surrounded with chitosan (AuNPs- chitosan).
  • an exemplary chitosan layer may have a thickness in a range of 10 nm to 100 nm.
  • an exemplary at least a stabilizing agent may stabilize an exemplary plurality of core- shell structured microbubbles in an exemplary PVA solution.
  • an exemplary plurality of AuNPs- chitosan may stabilize an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution by inhibiting gas diffusion, and reducing interfacial tension of an exemplary shell, which may also limit Ostwald ripening.
  • Ostwald ripening may refer to growth of one emulsion droplet at expense of a smaller one as a result of difference in chemical potential of material within exemplary droplets.
  • polyoxyethylene (20) sorbitan monooleate may stabilize an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution by reducing sedimentation of an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution via enhancing viscosity of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution.
  • an exemplary at least a fluorescent label may be added to PLGA-HP-PEG-cRGD-platelet.
  • an exemplary fluorescent label may include at least one of fluorescein isothiocyanate (FITC), rhodamine, and combinations thereof.
  • FITC may be added to PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 10:0.1 to 10:1 (PLGA-HP-PEG-cRGD-platelet: FITC).
  • FITC may have an excitation and an emission spectrum peak wavelengths of approximately 495 nm and 519 nm, respectively.
  • FITC may have a green color fluorescence emission.
  • an exemplary drug delivery composition may include an emulsion of an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution.
  • an exemplary PVA solution may include an aqueous PVA solution with a concentration in a range of 0.1 w/w% to 5 w/w%.
  • an exemplary emulsion of an exemplary plurality of core- shell structured microbubbles in an exemplary PVA solution may include a weight ratio in a range of 10:1 to 2:1 (an exemplary plurality of core-shell structured microbubbles: PVA).
  • an exemplary plurality of core- shell structured microbubbles may be stable in an exemplary PVA solution for a time period in a range of 1 day to 60 days.
  • a predetermined amount of an exemplary drug delivery composition may be injected intravenously to a patient suffering from MI.
  • an exemplary predetermined amount of an exemplary drug delivery composition may be in a range of 10 pL to 500 pL.
  • an exemplary drug delivery composition may reach an exemplary target place in an exemplary body of an exemplary patient.
  • cRGD may attach to GP Ub/IIIa receptors on activated platelet in arteries.
  • exemplary activated platelet may be accumulated inside arteries of an exemplary patient suffering from MI.
  • an exemplary target place may include platelet-clogged arteries that supply blood to heart.
  • an exemplary plurality of core-shell structured microbubbles may be systemically transferred using targeting ligands to an exemplary target place.
  • exemplary targeting ligands may include cRGD and platelet.
  • covalent or noncovalent attachment of exemplary targeting ligands on an exemplary shell may help recognizing antigens or receptors in an exemplary target place.
  • a fluorescent detector may be used to track FITC in an exemplary body of an exemplary patient.
  • an exemplary fluorescence detector may monitor fluorescent emission of an exemplary fluorescent label.
  • an exemplary fluorescent detector may be placed on chest of an exemplary patient.
  • an exemplary fluorescent detector may be placed on chest of an exemplary patent with a distance of an exemplary fluorescent detector to chest of an exemplary patient in a range of 1 cm to 5 cm h.
  • an exemplary fluorescent emission of an exemplary fluorescent label may be monitored utilizing an exemplary fluorescent detector for 20 hours to 72 hours.
  • tracking an exemplary fluorescent label may help to realize reaching an exemplary fluorescent label attached to PLGA-HP-PEG-cRGD-platelet to an exemplary target place.
  • an exemplary LIFU energy may be applied utilizing an ultrasonic device to a chest of an exemplary patient.
  • an exemplary LIFU energy may be applied utilizing an ultrasonic device with a distance to a chest of a patient in a range of 1 cm to 5 cm.
  • an exemplary an exemplary LIFU energy may be applied on a chest of an exemplary patient for a time period in a range of 1 minute to 30 minutes.
  • an exemplary LIFU energy may include acoustic intensity in a range of 1 mWcm” 2 to 600 mWcm” 2 .
  • an exemplary LIFU energy may include an acoustic frequency in a range of 1 MHz to 2 MHz.
  • an exemplary LIFU energy may induce an exemplary liquid PFH to evaporate.
  • evaporating an exemplary liquid PFH may increase internal pressure of an exemplary plurality of core-shell structured microbubbles.
  • an exemplary plurality of core-shell structures microbubbles may explode due to an exemplary increase of internal pressure of an exemplary plurality of core- shell structured microbubbles. Therefore, in an exemplary embodiment, bFGF may be released in an exemplary target place.
  • bFGF may induce blood vessel formation, promote proliferation restore perfusion and help migration of endothelial cells to infarcted area.
  • FIG. 1A illustrates a flowchart of a method 100 for healing myocardial infarction (MI), consistent with one or more exemplary embodiments of the present disclosure.
  • method 100 may include a step 102 of forming an emulsion of a plurality of coreshell structured microbubbles, step 104 of injecting a predetermined amount of the emulsion of the plurality of core-shell structured microbubbles intravenously to a body of a patient, step 106 of tracking a fluorescent label of each respective core- shell structured microbubble, and step 108 of applying an ultrasonic energy to chest of the patient.
  • step 102 of forming an emulsion of a plurality of core-shell structured microbubbles may include producing an exemplary plurality of coreshell structured microbubbles.
  • FIG. IB illustrates a flowchart of a method 110 to produce an exemplary plurality of core-shell structured microbubbles, consistent with one or more exemplary embodiments of the present disclosure.
  • method 110 may include a step 102a of forming a shell portion of each core-shell structured microbubble of a plurality of core- shell structured microbubbles, and a step 102b of forming a core portion of each core- shell structured microbubble of the plurality of core- shell structured microbubbles.
  • step 102a of forming an exemplary shell portion of each core-shell structured microbubble may include forming poly(D,L-lactide-co- glycolide)-NHS (PLGA-NHS) by dissolving PLGA-COOH, l-ethyl-3-(3- dimethylaminopropyl) carbodiimide (EDC), and NHS in an anhydrous dichloromethane solution.
  • PLGA-COOH may be mixed with EDC in a weight ratio in a range of 50: 1 to 10:1 (PLGA-COOH: EDC).
  • PLGA-COOH may be mixed with NHS in a weight ratio in a range of 50:1 to 5:1 (PLGA-COOH: NHS).
  • PLGA-COOH, EDC, and NHS may be mixed with an exemplary anhydrous dichloromethane solution with a weight ratio in a range of 1:1 to 1:2 (PLGA-COOH: an exemplary anhydrous dichloromethane solution).
  • PLGA-COOH, EDC, and NHS may be mixed with an exemplary anhydrous dichloromethane solution at a temperature in a range of 25°C to 35°C.
  • PLGA-COOH, EDC, and NHS may be mixed with an exemplary anhydrous dichloromethane solution under inert gas atmosphere.
  • an exemplary inert gas may include nitrogen.
  • FIG. 2A illustrates a chemical reaction of synthesizing PLGA-NHS, consistent with one or more exemplary embodiments of the present disclosure.
  • step 102a may further include forming PLGA-NH2 by dissolving formed PLGA-NHS and ethylenediamine (EDA) in a dichloromethane solution.
  • EDA ethylenediamine
  • PLGA-NHS and EDA may be dissolved in an exemplary dichloromethane solution with a weight ratio in a range of 10:1 to 10:1.2 (PLGA-NHS: EDA).
  • PLGA-NHS and EDA may be dissolved in an exemplary dichloromethane solution at a temperature in a range of 25°C to 35°C.
  • PLGA-NHS and EDA may be mixed in an exemplary dichloromethane solution using a mixer with a rotational speed in a range of 100 rpm to 500 rpm.
  • PLGA-NHS and EDA may be mixed in an exemplary dichloromethane solution for 60 minutes to 2000 minutes.
  • FIG 2B illustrates a chemical reaction of synthesizing PLGA-NH2, consistent with one or more exemplary embodiments of the present disclosure.
  • step 102a may further include forming PLGA-HP by mixing dissolved heparin salt, NHS, and EDC in N,N- dimethylformamide/formamide (DMF/FA) with an exemplary formed PLGA-NH2.
  • DMF/FA N,N- dimethylformamide/formamide
  • a chemical reaction between carboxyl group of activated heparin and amine group of PLGA may happen.
  • an exemplary chemical reaction may lead to amide bond formation between PLGA and heparin.
  • DMF and FA may be mixed with a weight ratio in a range of 10:1 to 1:1 (DMF: FA).
  • heparin salt, NHS, EDC, and an exemplary formed PLGA-NH2 may be mixed in DMF/FA with a weight ratio in a range of 1:1 to 1:5 (heparin: NHS).
  • heparin salt, NHS, EDC, and an exemplary formed PLGA-NH2 may be mixed in DMF/FA with a weight ratio in a range of 10:1 to 1:1 (heparin: EDC).
  • heparin salt, NHS, EDC, and an exemplary formed PLGA-NH2 may be mixed in DMF/FA with a weight ratio in a range of 1:1 to 1:5 (heparin: an exemplary formed PLGA-NH2).
  • heparin salt, NHS, EDC, and an exemplary formed PLGA-NH2 may be mixed in DMF/FA at a temperature in a range of 25°C to 35°C.
  • FIG 2C illustrates a chemical reaction of synthesizing PEGA-HP, consistent with one or more exemplary embodiments of the present disclosure
  • step 102a may further include forming HOOC-PEG-COOH by dissolving HO-PEG-OH, succinic anhydride, and 4- (dimethylamino) pyridine (DMAP) in a dichloromethane solution.
  • HO-PEG-OH, succinic anhydride, and DMAP may be mixed in an exemplary dichloromethane solution utilizing a mixer with a rotational speed in a range of 100 rpm to 1000 rpm for time period in a range of 24 hours to 50 hours.
  • HO-PEG-OH, succinic anhydride, and DMAP may be mixed in an exemplary dichloromethane solution with a weight ratio in a range of 20:1 to 10:1 (HO-PEG-OH: DMAP).
  • HO-PEG-OH, succinic anhydride, and DMAP may be mixed in an exemplary dichloromethane solution with a weight ratio in a range of 2: 1 to 1 : 1 (HO-PEG-OH: succinic anhydride).
  • step 102a may further include forming HOOC-PEG-NHS by dissolving an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an anhydrous dichloromethane solution.
  • dissolving an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an anhydrous dichloromethane solution may include mixing an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an exemplary anhydrous dichloromethane solution utilizing a mixer with a rotational speed in a range of 100 rpm to 1000 rpm.
  • dissolving an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an exemplary anhydrous dichloromethane solution may include mixing an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an exemplary anhydrous dichloromethane solution utilizing an exemplary mixer for a time period in a range of 15 hours to 50 hours.
  • dissolving an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an anhydrous dichloromethane solution may be performed under an inert gas atmosphere to activate carboxylic groups of PEG.
  • an exemplary inert gas may include nitrogen.
  • step 102a may further include forming PLGA-HP-PEG-NHS by mixing an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP and dicyclohexylcarbodiimide (DCC).
  • DCC dicyclohexylcarbodiimide
  • an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP may be mixed with dicyclohexylcarbodiimide (DCC) utilizing an exempalry mixer for a time period in a range of 20 hours to 30 hours.
  • DCC dicyclohexylcarbodiimide
  • an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP may be mixed with DCC under an inert gas atmosphere.
  • an exemplary inert gas may include nitrogen.
  • an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP may be mixed with DCC with a weight ratio in a range of 1:1 to 1:10 (HOOC-PEG-NHS: PLGA-HP).
  • an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP may be mixed with DCC with a weight ratio in a range of 100:1 to 10:1 (HOOC-PEG-NHS: DCC).
  • step 102a may further include forming PLGA-HP-PEG-cRGD by mixing an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA with a cyclic arginine-glycine-aspartate (cRGD) solution.
  • cRGD cyclic arginine-glycine-aspartate
  • an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA may be added dropwise to an exemplary cRGD solution.
  • an exemplary cRGD solution may have a concentration in a range of 1 mol/L to 100 mol/L.
  • an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA may be mixed with an exemplary cRGD solution utilizing a mixer with a rotational speed in a range of 1 rpm to 10 rpm.
  • an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA may be mixed with an exemplary cRGD solution utilizing an exemplary mixer for a time period in a range of 200 minutes to 2000 minutes.
  • FIG 2D illustrates a chemical reaction for synthesizing PLGA-HP-PEG-cRGD, consistent with one or more exemplary embodiments of the present disclosure.
  • step 102a may further include forming PLGA-HP-PEG-cRGD-platelet by mixing a platelet solution with an exemplary formed PLGA-HP-PEG-cRGD.
  • platelet may be separated from blood by differential centrifugation.
  • blood may be centrifuged with a rotational speed in a range of 2000 rpm to 4000 rpm for a time period in a range of 10 minutes to 30 minutes.
  • an exemplary platelet solution may be mixed with an exemplary formed PLGA-HP-PEG-cRGD utilizing a mixer with a rotational speed in a range of 100 rpm to 500 rpm.
  • an exemplary platelet solution may be mixed with an exemplary formed PLGA-HP-PEG-cRGD utilizing an exemplary mixer for a time period in a range of 60 minutes to 2000 minutes.
  • an exemplary platelet solution may be mixed with an exemplary formed PLGA-HP-PEG-cRGD with a weight ratio in a range of 100:1 to 10:1 (PLGA-HP- PEG-cRGD: platelet).
  • step 102a may further include forming bFGF loaded PLGA-HP-PEG-cRGD-platelet by mixing an exemplary formed PLGA-HP-PEG-cRGD-platelet with bFGF dissolved in a phosphate-buffered saline (PBS) solution.
  • PBS phosphate-buffered saline
  • an exemplary formed PLGA-HP-PEG-cRGD-platelet may be mixed with an exemplary bFGF solution utilizing a mixer with a rotational speed in a range of 100 rpm to 500 rpm.
  • an exemplary formed PLGA-HP-PEG-cRGD-platelet may be mixed with an exemplary bFGF solution for a time period in a range of 100 minutes to 3000 minutes.
  • an exemplary PBS solution may have a concentration in a range of 1 mol/L to 10 mol/L.
  • an exemplary bFGF solution may have a concentration in a range of 1 mol/L to 100 mol/L.
  • an exemplary formed PLGA- HP-PEG-cRGD-platelet may be mixed with an exemplary bFGF solution with a weight ratio in a range of 10:1 to 2:1 (an exemplary formed PLGA-HP-PEG-cRGD-platelet: bFGF).
  • step 102a may further include forming a plurality of fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet structures.
  • forming an exemplary plurality of fluorescent labeled bFGF loaded PLGA-HP- PEG-cRGD-platelet structures may include mixing at least a fluorescent label with bFGF loaded PLGA-HP-PEG-cRGD-platelet.
  • an exemplary at least a fluorescent label may include at least one of fluorescein isothiocyanate (FITC), rhodamine, and combinations thereof.
  • FITC may be attached to bFGF.
  • FITC may be mixed with an exemplary bFGF loaded PLGA-HP-PEG- cRGD-platelet structures with a weight ratio in a range of 1:10 to 1:100 (FITC: an exemplary plurality of bFGF loaded PLGA-HP-PEG-cRGD-platelet structures).
  • step 102a may further include adding at least a stabilizing agent to bFGF loaded PLGA-HP-PEG-cRGD-platelet.
  • an exemplary at least a stabilizing agent may be mixed with bFGF loaded PLGA- HP-PEG-cRGD-platelet utilizing a mixer with a rotational speed in a range of 100 rpm to 1000 rpm.
  • an exemplary at least a stabilizing agent may be mixed with bFGF loaded PLGA-HP-PEG-cRGD-platelet utilizing an exemplary mixer for a time period in a range of 1200 minutes to 2500 minutes.
  • an exemplary stabilizing agent may include at least one of a plurality of gold nanoparticles (AuNPs), polyoxyethylene (20) sorbitan monooleate, and combinations thereof.
  • AuNPs gold nanoparticles
  • polyoxyethylene (20) sorbitan monooleate polyoxyethylene (20) sorbitan monooleate
  • an exemplary plurality of AuNPs may be synthesized using chitosan as a reducing agent.
  • an exemplary chitosan may be wrapped over an exemplary plurality of AuNPs (AuNPs-chitosan).
  • an exemplary plurality of AuNPs may be synthesized by heating an aqueous solution of HAuCL and a chitosan solution at a temperature in a range of 40°C to 100°C for a time period in a range of 30 minutes to 300 minutes.
  • each respective AuNP-chitosan may include an AuNP surrounded with a layer of chitosan with a thickness in a range of 10 nm to 100 nm.
  • an exemplary plurality of core- shell structured microbubbles may include polyoxyethylene (20) sorbitan monooleate with a weight ratio in a range of 1:10 to 1:1 (polyoxyethylene (20) sorbitan monooleate: PLGA-HP-PEG-cRGD- platelet).
  • an exemplary plurality of AuNPs may be attached to an exemplary shell of an exemplary plurality of core-shell structured microbubbles.
  • an exemplary plurality of core- shell structured microbubbles may include an exemplary plurality of AuNPs-chitosan with a weight ratio in a range of 1:1000 to 1:10 (Au NPs-chitosan: PLGA-HP-PEG-cRGD-platelet).
  • an exemplary plurality of core- shell structured microbubbles may include polyoxyethylene (20) sorbitan monooleate with a weight ratio in a range of 1 : 10 to 1:1 (polyoxyethylene (20) sorbitan monooleate: PLGA-HP-PEG-cRGD-platelet).
  • step 102a may further include fluorescent labeling an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet with at least a fluorescent label.
  • an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet may be fluorescent labeled by mixing an exemplary at least a fluorescent label with bFGF loaded PLGA-HP-PEG-cRGD-platelet.
  • an exemplary fluorescent label may include at least one of fluorescein isothiocyanate (FITC), rhodamine, and combinations thereof.
  • FITC may attach to bFGF.
  • a FITC solution in dimethyl sulfoxide (DMSO) may be added to an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:20 to 1:10 (FITC: bFGF loaded PLGA-HP-PEG-cRGD-platelet).
  • a FITC solution and an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet may be mixed for a time period in a range of 60 minutes to 1800 minutes.
  • an exemplary FITC solution and an exemplary bFGF loaded PLGA-HP-PEG- cRGD-platelet may be mixed in darkness.
  • step 102a may further include injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution into a PVA solution using a first syringe pump.
  • an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution may be injected into a microfluidic device.
  • an exemplary PVA solution may be injected into an exemplary microfluidic device before injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution.
  • injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution may include injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG- cRGD-platelet solution into an exemplary PVA solution using an exemplary first syringe pump with a flow rate in a range of 100 pl/min to 500 pl/min.
  • an exemplary PVA solution may have a concentration in a range of 0.1 w/w% to 5 w/w% in water.
  • an exemplary solution of bFGF loaded PLGA-HP-PEG-cRGD- platelet in phosphate-buffered saline (PBS) may be injected into a microfluidic device utilizing an exemplary first syringe pump.
  • step 102b of forming a core portion of each core- shell structured microbubble may include injecting a liquid perfluorohexane (PFH) into an exemplary PVA solution.
  • PFH liquid perfluorohexane
  • an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution and an exemplary liquid PFH may be injected into an exemplary PVA solution, simultaneously.
  • an exemplary liquid PFH may be injected into an exemplary microfluidic device utilizing a second syringe pump.
  • an exemplary liquid PFH may be injected into an exemplary PVA solution with a flow rate in a range of 1 pl/min to 500 pl/min.
  • an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution and an exemplary liquid PFH may be injected into an exemplary PVA solution with a weight ratio in a range of 1:10 to 1:1 (bFGF loaded PLGA-HP-PEG-cRGD-platelet solution: liquid PFH).
  • an exemplary PVA solution may have a concentration in a range of 0.1 w/w% to 5 w/w% in water.
  • step 102 of forming an emulsion of a plurality of core- shell structured microbubbles may include stabilizing an exemplary emulsion of a plurality of core- shell structured microbubbles in a PVA solution.
  • each respective core-shell structured microbubble of an exemplary plurality of core-shell structured microbubbles may have a size in a range of 1 pm to 30 pm.
  • an exemplary emulsion of a plurality of core-shell structured microbubbles may be stabilized sing at least a stabilizing agent.
  • an exemplary at least a stabilizing agent may include at least one of a plurality of gold nanoparticles (AuNPs), polyoxyethylene (20) sorbitan monooleate, and combinations thereof.
  • AuNPs gold nanoparticles
  • polyoxyethylene (20) sorbitan monooleate may be attached to an exemplary shell of an exemplary plurality of core- shell structured microbubbles with a weight ratio in a range of 1:10 to 1:1 (polyoxyethylene (20) sorbitan monooleate: an exemplary shell).
  • polyoxyethylene (20) sorbitan monooleate may be physically attached on an exemplary shell.
  • an exemplary plurality of AuNPs may be attached to an exemplary shell of an exemplary plurality of coreshell structured microbubbles.
  • an exemplary plurality of AuNPs may stabilize an exemplary emulsion by inhibition of gas diffusion to an exemplary plurality of core- shell structured microbubbles due to presence of solid AuNPs.
  • an exemplary plurality of AuNPs may stabilize an exemplary emulsion by reducing interfacial tension which may also limit Ostwald ripening.
  • an exemplary plurality of AuNPs may be produced using chitosan as a reducing agent.
  • each respective AuNP may include a layer of chitosan wrapped over an exemplary AuNP (AuNP-chitosan).
  • an exemplary layer of chitosan may have a thickness in a range of 10 nm to 100 nm.
  • AuNPs- chitosan may be produced by mixing an aqueous solution of HAuCh with an aqueous solution of chitosan in a mixer with a rotational speed in a range of 100 rpm to 1000 rpm.
  • an exemplary solution of HAuCU may be added to an exemplary aqueous solution of chitosan with a weight ratio in a range of 1:100 to 1:10 (chitosan: HAuCU).
  • exemplary AuNPs-chitosan may be produced at a temperature in a range of 30°C to 100°C.
  • an exemplary plurality of AuNPs-chitosan may be attached to PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:1000 to 1:10 (Au NPs-chitosan: PLGA-HP-PEG-cRGD- platelet).
  • each respective AuNP of an exemplary plurality of AuNPs may have a diameter in a range of 25 nm to 75 nm.
  • PVA may also stabilize an exemplary emulsion by reducing sedimentation of an exemplary plurality of core-shell structured microbubbles via enhancing viscosity of an exemplary emulsion.
  • FIG. 3 illustrates a microfluidic device 300 for producing an exemplary plurality of core- shell structured microbubbles, consistent with one or more exemplary embodiments of the present disclosure.
  • microfluidic device 300 may include a plate 312, a microfluidic vessel 310, a first inlet 302, a second inlet 304, a third inlet 306, and an outlet 308.
  • plate 312 may be made of at least one of polydimethyl siloxane (PDMS), glass, or polymethyl methacrylate (PMMA).
  • PDMS polydimethyl siloxane
  • PMMA polymethyl methacrylate
  • plate 312 may be a platform for holding microfluidic vessel 310, first inlet 302, second inlet 304, third inlet 306, and outlet 308 thereon.
  • first inlet 302 may be used for injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP- PEG-cRGD-platelet solution into microfluidic vessel 310 utilizing an exemplary first syringe pump.
  • second inlet 304 may be used for injecting an exemplary liquid PFH into microfluidic vessel 310 utilizing an exemplary second syringe pump.
  • an exemplary PVA solution may be injected via third inlet 306 into microfluidic vessel 310.
  • outlet 308 may be used to collect an exemplary drug delivery composition produced by method 100 and method 110.
  • an exemplary drug delivery composition may be an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution.
  • an exemplary PVA solution may have a concentration in a range of 0.1 to 5 wlw c /c in water.
  • each respective core- shell structured microbubble of an exemplary plurality of core- shell structured microbubbles may have a size in a range of 1 pm to 100 pm.
  • step 104 of injecting a predetermined amount of an exemplary drug delivery composition may include injecting an exemplary predetermined amount of an exemplary drug delivery composition intravenously to a body of a patient utilizing an ampule.
  • injecting an exemplary predetermined amount of an exemplary drug delivery composition may include injecting an amount of an exemplary drug delivery composition in a range of 10 pL to 500 pL intravenously to an exemplary body of an exemplary patient.
  • an exemplary drug delivery composition may be injected intravenously to a body of a patient after a time period in a range of 60 minutes to 7200 minutes after producing an exemplary drug delivery composition due to coagulation of an exemplary plurality of core-shell structured microbubbles.
  • an exemplary drug delivery composition may be injected intravenously to a body of a human and a body of an animal.
  • step 106 of tracking a fluorescent label of each respective core-shell structured microbubble may include monitoring an exemplary fluorescent label of each respective core-shell structured microbubble.
  • tracking an exemplary fluorescent label of each respective core-shell structured microbubble may include tracking an exemplary fluorescent label of each respective core-shell structured microbubble utilizing a fluorescent detector.
  • an exemplary drug delivery composition may include at least a fluorescent label.
  • an exemplary fluorescent label may include at least one of FITC, rhodamine, and combinations thereof.
  • an exemplary fluorescent label attached to bFGF in an exemplary drug delivery composition may be monitored utilizing an exemplary fluorescent detector after injecting an exemplary drug delivery composition intravenously to a body of an exemplary patient.
  • bFGF may be attached to FITC for monitoring in vivo migration of an exemplary plurality of core- shell structured microbubbles through blood vessels.
  • FITC may emit light with a wavelength in a green region.
  • an exemplary fluorescent detector may show existence of an exemplary core-shell structured microbubbles using fluorescence emission of FITC.
  • an exemplary fluorescence detector may be placed on chest of an exemplary patient.
  • accumulation of FITC in an exemplary target place may be understood when an exemplary target place turns to a green color (shown by an exemplary fluorescent detector).
  • FITC may be attached to bFGF of an exemplary drug delivery composition. Therefore, green color emission in an exemplary target place may show existence of an exemplary drug delivery composition in an exemplary target place.
  • an ultrasonic device may be placed on chest of an exemplary patient.
  • step 108 of applying an ultrasonic energy to chest of an exemplary patient may include applying an exemplary ultrasonic energy in a distance in a range of 1 cm to 5 cm to chest of an exemplary patient.
  • an exemplary ultrasonic energy may be applied on chest of an exemplary patient.
  • an exemplary drug delivery composition may reach to an exemplary target place in a time period of maximum 72 hours.
  • an exemplary ultrasonic energy may be an acoustic energy with acoustic frequency in a range of 1 MHz to 2 MHz.
  • an exemplary ultrasonic energy may be applied utilizing an ultrasonic device with acoustic intensity in a range of 1 mWcm” 2 to 600 mWcm" 2 .
  • applying an exemplary ultrasonic energy may include applying an exemplary ultrasonic energy on an exemplary chest of an exemplary patient for a time period in a range of 1 minute to 30 minutes.
  • applying an exemplary ultrasonic energy to chest of an exemplary patient may include applying an exemplary ultrasonic energy to chest of an exemplary patient after reaching an exemplary drug delivery composition to an exemplary target place.
  • an exemplary target place may include platelet-clogged arteries that supply blood to heart.
  • an exemplary liquid PFH may vaporize.
  • an exemplary plurality of core-shell structured micro bubbles may get larger with an exemplary vaporization process.
  • an exemplary plurality of core-shell structured micro bubbles may expand until an exemplary core-shell structured micro bubbles may lose an exemplary core-shell structure and may explode.
  • an exemplary plurality of core-shell structured micro bubbles may lose an exemplary core-shell structure after a time period in a range of 1 minute to 120 minutes of ultrasonic illumination.
  • bFGF after exploding an exemplary plurality of core-shell structured microbubbles, bFGF may be released in an exemplary target place.
  • an exemplary slow process of explosion may help to controlled release of bFGF.
  • all of an exemplary plurality of core-shell structured micro bubbles may explode.
  • bFGF may help blood vessel formation, promote proliferation restore perfusion and help migration of endothelial cells in infarcted area.
  • Example 1 Producing PLGA-HP-PEG-cRGD-platelet
  • PLGA-HP For producing PLGA-HP, a method similar to step 102 of method 100 and/or method 110 may be used.
  • low molecular weight heparin salt (0.02 mmol), NHS (0.27 mmol), and EDC (0.05 mmol) were dissolved in DMF/FA (5 mL).
  • PLGA-NH2 (0.004 mmol) was reacted at room temperature. Then, the product was dialyzed against distilled water for three days. The product was then lyophilized to obtain PLGA-HP.
  • PLGA-HP conjugated HOOC-PEG-NHS was obtained by a direct coupling reaction.
  • PLGA-HP (0.005 mmol) and HOOC-PEG-NHS (0.01 mmol) were dissolved in the mixture of DMF/FA solvent. Then, DMAP and DCC were added to the solution. The mixture of PLGA-HP, HOOC-PEG-NHS, DMAP and DCC (reaction mixture) were mixed.
  • the reaction mixture was precipitated in cold diethyl ether after the mixing process.
  • a solution of PLGA-HP-PEG-NHS in anhydrous DMF/FA was poured dropwise into the cRGD solution. After stirring, the obtained solution was dialyzed to remove the unreacted mixture.
  • the conjugate PLGA-HP-PEG-cRGD was then lyophilized.
  • platelets were separated from whole sheep blood by differential centrifugation.
  • the plasma blood bag was poured into a falcon and centrifuged to remove RBC.
  • platelet-rich plasma (PRP) was prepared by adding blood plasma into a tube and centrifuging the blood plasma. Platelets were re-suspended in PBS using anticoagulant ACD. Then, the mixture of platelets and PBS were added to PLGA-HP-PEG-cRGD.
  • Example 2 Producing microbubbles of bFGF-loaded PLGA-HP-PEG-cRGD- [0073]
  • a method similar to method 100 and method 110 was used.
  • PLGA-HP-PEG-cRGD (50 mg) and bFGF (50 pg) were suspended in PBS.
  • bFGF-loaded PLGA-HP-PEG-cRGD was collected by centrifugation and washed with PBS to remove free bFGF.
  • bFGF- loaded PLGA-HP-PEG-cRGD 300 pL
  • gold nanoparticles 300 pL
  • polyoxyethylene (20) sorbitan monooleate was added to the mixture.
  • the obtained solution and perfluorohexane were injected using two independent syringe pumps at a predetermined constant flow rate.
  • the obtained droplets from the microfluidic device were stabilized in a PVA solution and collected.
  • Microfluidic devices allow controlling the flow rates of two different solutions by employing two independent pumps.
  • the inner solution was a perfluorohexane (PFH).
  • PFH perfluorohexane
  • the outer aqueous phase was the polymeric solution containing PLGA-HP-PEG-cRGD/gold nanoparticles/ polyoxyethylene (20) sorbitan monooleate mixture.
  • Droplets were made by using a microfluidic device demonstrated in FIG. 3. The final droplets were collected in a PVA solution as a stabilizing agent.
  • FIG. 4 illustrates a spectrum of proton nuclear magnetic resonance (1HNMR) spectroscopy of PLGA-HP-PEG-NHS, consistent with one or more exemplary embodiments of the present disclosure.
  • the 1H-NMR spectrum of the final product displayed successful synthesis of PLGA-HP-PEG copolymer in which the main signals were related to PLGA moiety (a), (b), (c) at 1.46, 4.8, and 5.2 ppm of the lactide units in PLGA, respectively.
  • the chemical shifts at 3.38 and 4.1 ppm (d, e) were related to the NH-CH2-CH2-NH, respectively.
  • the chemical shifts at 2.1 ppm (f), 3.1, and 3.5 ppm (g, h) were indicatives of CH-NH of heparin and PEG moiety (methylene hydrogen of the PEG), respectively (FIG. 4).
  • FIG. 5 illustrates an optical microscopy image of microbubbles (MBs) obtained at flow rates of 1 pl/min for PFH and 250 pl/min for an exemplary bFGF loaded PLGA-HP-PEG- cRGD-platelet solution, consistent with one or more exemplary embodiments of the present disclosure.
  • PFH-droplets are interesting for ultrasound-triggered delivery systems. To disrupt the PFH-loaded microspheres were exposed to low intensity focused ultrasound (LIFU) (an acoustic frequency of 1.5 MHz and an acoustic intensity of 35 mW/cm 2 ). When exposing the droplets to LIFU, the variations of the formed micro bubbles (MBs) were observed via light microscopy.
  • LIFU low intensity focused ultrasound
  • FIG. 6A illustrates an image of MBs before applying LIFU, consistent with one or more exemplary embodiments of the present disclosure.
  • FIG. 6B illustrates an image of MBs after 20 minutes of applying LIFU with acoustic intensity of 35 mW/cm 2 , consistent with one or more exemplary embodiments of the present disclosure.
  • FIG. 6C illustrates an image of MBs after 45 minutes of applying LIFU with acoustic intensity of 35 mW/cm 2 , consistent with one or more exemplary embodiments of the present disclosure.
  • FIG. 6A illustrates an image of MBs before applying LIFU, consistent with one or more exemplary embodiments of the present disclosure.
  • FIG. 6B illustrates an image of MBs after 20 minutes of applying LIFU with acoustic intensity of 35 mW/cm 2 , consistent with one or more exemplary embodiments of the present disclosure.
  • FIG. 6C illustrates an image of MBs after 45 minutes of applying LIFU with acoustic intensity of 35
  • FIG. 6D illustrates an image of MBs after 7 minutes of applying LIFU with acoustic intensity of 330 mW/cm 2 , consistent with one or more exemplary embodiments of the present disclosure.
  • FIG. 6E illustrates an image of MBs after 12 minutes of applying LIFU with acoustic intensity of 330 mW/cm 2 , consistent with one or more exemplary embodiments of the present disclosure.
  • FIG. 7 illustrates fluorescence microscopic images of drug delivery samples for analyzing cellular uptake, consistent with one or more exemplary embodiments of the present disclosure.
  • UTS stands for ultrasound irradiation.
  • different samples were tested with UTS and without UTS.
  • control sample (702), bFGF sample (704), bFGF- PLGA-HP-PEG-cRGD sample (706), and bFGF- PLGA-HP-PEG-cRGD-platelet sample (708) were illuminated by an ultrasonic energy.
  • Control sample (710), bFGF sample (712), bFGF- PLGA-HP-PEG-cRGD sample (714), and bFGF- PLGA-HP-PEG-cRGD-platelet sample (716) were analyzed before an ultrasonic illumination.
  • Results showed cellular uptake of bFGF- PLGA-HP-PEG-cRGD- platelet samples was higher due to interaction between platelet and activated platelet in blood vessels.
  • Example 3 Analyzing in-vivo targeting ability of an exemplary drug delivery
  • FIG. 8 illustrates fluorescence microscopic images of a plurality of fluorescent labeled core- shell structured microbubbles (804) and a plurality of core-shell structured microbubbles with no label (802), consistent with one or more exemplary embodiments of the present disclosure.
  • the plurality of fluorescent labeled core- shell structured microbubbles had fluorescent images with green color indicating the presence of the plurality of core-shell structured microbubbles.
  • the present disclosure may include a drug delivery composition for healing MI.
  • An exemplary drug delivery composition may include an emulsion of a plurality core- shell structured microbubble in a PVA solution.
  • An exemplary emulsion may be injected intravenously to a patient.
  • An exemplary drug delivery composition may be activated by an ultrasonic energy applied to chest of an exemplary patient.
  • An exemplary drug delivery composition may be used in hospitals and clinics to treat MI and hinder side effects of oxygen deficiency of heart.

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Abstract

A drug delivery composition for healing myocardial infarction (MI) is disclosed. The drug delivery composition includes a plurality of core-shell structured microbubbles and a polyvinyl alcohol (PVA) solution. The plurality of core-shell structured microbubbles is dispersed in the PVA solution. Each core-shell structured microbubble of the plurality of core-shell structured microbubbles includes a core and a shell surrounding the core. The core includes liquid perfluorohexane (PFH). The shell includes poly(D,L-lactide-co-glycolide)-heparin-polyethylene glycol-cyclic arginine glycine aspartate-platelet (PLGA-HP-PEG-cRGD- platelet), basic fibroblast growth factor (bFGF), at least one stabilizing agent, and at least a fluorescent label.

Description

A DRUG DELIVERY COMPOSITION FOR TREATING MYOCARDIAL
INFRACTION (MI)
TECHNICAL FIELD
[0001] The present disclosure generally relates to a drug delivery composition for healing myocardial infarction (MI), and more particularly, relates to a method of producing and administering a drug delivery composition for healing MI.
BACKGROUND ART
[0002] Cardiovascular disease has been known as one of a leading cause of death. Myocardial infarction (MI) is a common cardiovascular disease which can cause an irreversible death of heart muscle due to lack of oxygen. MI undergoes progressive conditions that usually leads to myocardial fibrosis, diastolic function, and even weakened ventricular contraction, heart failure (HF), malignant arrhythmia, and sudden death.
[0003] One of methods used for healing MI is direct injection of basic fibroblast growth factor (bFGF) into heart. Direct injection of bFGF into heart has many drawbacks, i.e., health risks which can limit its clinical application. As a result, different drug delivery systems have been introduced such as systematic drug delivery. Systemic drug delivery is a method of administering a drug, a nutrition, etc., so that whole body is affected. Systemic drug delivery has low efficiency because of slow tissue penetration, short in-vivo half-life, and tendency to cause systemic side effects.
[0004] Different strategies have been applied in prior art for healing MI. For example, Mark A. et al. presented a patent on “Systems, methods, and devices for production of gas-filled microbubbles” (WO-2014066624-A1). Mark A. et al. used a continuous flow chamber and a sonicator to synthesize gas-filled microbubbles. A microbubble solution produced by the introduced method and device can be size-sorted for a particular application, such as injection into a patient for gas delivery thereto. Eleanor Stride et al. presented a patent on “Method and apparatus for generating bubbles” (US 11007495 B2). Eleanor Stride et al. used a method to produce bubbles by injecting a stream of the first fluid into the microfluidic channel through an aperture such that bubbles of the first fluid form in the second fluid. Jon Nagy et al. presented a patent on “Polymerized shell lipid microbubbles and uses thereof’ (US 20130129635 Al). Jon Nagy et al. produced shell lipid microbubbles (PSMs) for diagnosis and treatment of a condition. Microbubbles produced by these methods may suffer from short life time, low stability, and bio-incompatibility.
[0005] There is, therefore, a need for a cost-effective, biocompatible, high efficiency drug delivery composition for treating MI being capable of accurately transferring drugs to a target place. There is further a need for a fast method to produce and administer a drug delivery composition for treating MI.
SUMMARY OF THE DISCLOSURE
[0006] This summary is intended to provide an overview of the subject matter of this patent, and is not intended to identify essential elements or key elements of the subject matter, nor is it intended to be used to determine the scope of the claimed implementations. The proper scope of this patent may be ascertained from the claims set forth below in view of the detailed description below and the drawings.
[0007] According to one or more exemplary embodiments, the present disclosure is directed to a drug delivery composition for healing myocardial infarction (MI). In an exemplary embodiment, an exemplary drug delivery composition may include a plurality of core-shell structured microbubbles and a polyvinyl alcohol (PVA) solution. In an exemplary embodiment, each respective core- shell structured microbubble may include a core and a shell surrounding an exemplary core. In an exemplary embodiment, an exemplary core may include liquid perfluorohexane (PFH). In an exemplary embodiment, an exemplary shell may include poly(D,L-lactide-co-glycolide)-heparin-polyethylene glycol-cyclic arginine glycine aspartate - platelet (PLGA-HP-PEG-cRGD- platelet), basic fibroblast growth factor (bFGF) attached to heparin of PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:10000 to 1:10 (bFGF: PLGA-HP-PEG-cRGD-platelet), at least one stabilizing agent attached to PLGA-HP- PEG-cRGD- platelet, and at least one fluorescent label attached to bFGF. In an exemplary embodiment, an exemplary liquid PFH to an exemplary PLGA-HP-PEG-cRGD-platelet may have a weight ratio in a range of 1:10 to 1:1 (liquid PFH: PLGA-HP-PEG-cRGD-platelet). In an exemplary embodiment, an exemplary plurality of core-shell structured microbubbles may be dispersed in an exemplary PVA solution.
[0008] In an exemplary embodiment, an exemplary at least one stabilizing agent may include at least one of a plurality of gold nanoparticles (AuNPs), polyoxyethylene (20) sorbitan monooleate, and combinations thereof. In an exemplary embodiment, an exemplary at least one stabilizing agent may include a plurality of AuNPs may be surrounded by chitosan (AuNPs-chitosan). In an exemplary embodiment, each respective AuNP-chitosan may include an AuNP surrounded with a layer of chitosan with a thickness in a range of 10 nm to 100 nm. [0009] In an exemplary embodiment, each respective AuNP of an exemplary plurality of AuNPs may have a particle size in a range of 25 nm to 75 nm. In an exemplary embodiment, an exemplary drug delivery composition may include an exemplary plurality of AuNPs- chitosan to an exemplary PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:1000 to 1:10 (Au NPs-chitosan: PLGA-HP-PEG-cRGD-platelet). In an exemplary embodiment, an exemplary drug delivery composition may include polyoxyethylene (20) sorbitan monooleate to an exemplary PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:10 to 1:1 (polyoxyethylene (20) sorbitan monooleate: PLGA-HP-PEG-cRGD- platelet).
[0010] In an exemplary embodiment, an exemplary PVA solution may include an aqueous PVA solution with a concentration of PVA in a range of 0.1 w/w% to 5 w/w%. In an exemplary embodiment, an exemplary at least one fluorescent label may include at least one of fluorescein isothiocyanate, rhodamine, and combinations thereof. In an exemplary embodiment, each respective core-shell structured microbubble of an exemplary plurality of core-shell structured microbubbles may have a size in a range of 1 pm to 30 pm.
[0011] According to one or more exemplary embodiments, the present disclosure is directed to a method for treating myocardial infarction (MI). In an exemplary embodiment, an exemplary method may include forming an emulsion of a plurality of core- shell structured microbubbles in which each respective core- shell structure microbubble may have a fluorescent label, injecting a predetermined amount of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles intravenously to a body of a patient, tracking a fluorescent label of each respective core-shell structure microbubble utilizing a fluorescent detector for a time period of maximum 72 hours, and applying an ultrasonic energy to chest of an exemplary patient with acoustic intensity in a range of 1 mWcm”2 to 600 mWcm"2.
[0012] In an exemplary embodiment, injecting an exemplary predetermined amount of an exemplary emulsion of an exemplary plurality of core- shell structured microbubbles intravenously to an exemplary body of an exemplary patient may include injecting an amount of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles in a range of 10 pg/mL to 500 pg/mL intravenously to the body of the patient. [0013] In an exemplary embodiment, applying an exemplary ultrasonic energy an exemplary chest of an exemplary patient may include applying an exemplary ultrasonic energy with an acoustic frequency in a range of 1 MHz to 2 MHz to an exemplary chest of an exemplary patient.
[0014] In an exemplary embodiment, applying an exemplary ultrasonic energy to an exemplary chest of an exemplary patient may include applying an exemplary ultrasonic energy to an exemplary chest of an exemplary patient after reaching an exemplary drug delivery composition to an exemplary target place, in which an exemplary target place may include platelet-clogged arteries that supply blood to heart.
[0015] In an exemplary embodiment, applying an exemplary ultrasonic energy may include applying an exemplary ultrasonic energy to an exemplary chest of an exemplary patient for a time period in a range of 1 minute to 30 minutes.
[0016] In an exemplary embodiment, forming an exemplary emulsion of an exemplary plurality of core- shell structured microbubbles may include forming a shell portion and forming a core portion. In an exemplary embodiment, forming an exemplary shell portion may include forming poly(D,L-lactide-co-glycolide)-NHS (PLGA-NHS) by dissolving PLGA-COOH, l-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), and NHS in an anhydrous dichloromethane solution, forming PLGA-NH2 by dissolving formed PLGA-NHS and ethylenediamine (EDA) in a dichloromethane solution, forming PLGA-HP by mixing dissolved heparin salt, NHS, and EDC with an exemplary formed PLGA-NH2in N,N- dimethylformamide/formamide (DMF/FA), forming HOOC-PEG-COOH by dissolving HO-PEG-OH, succinic anhydride, and 4-(dimethylamino) pyridine (DMAP) in a dichloromethane solution, forming HOOC-PEG-NHS by dissolving an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an anhydrous dichloromethane solution, forming PLGA-HP-PEG-NHS by mixing an exemplary formed HOOC-PEG-NHS, an exemplary formed PLGA-HP, DMAP, and dicyclohexylcarbodiimide (DCC), forming PLGA-HP-PEG- cRGD by mixing an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA with a cyclic arginine-glycine-aspartate (cRGD) solution, forming PLGA-HP-PEG-cRGD-platelet by mixing a platelet solution with an exemplary formed PLGA-HP-PEG-cRGD, forming bFGF loaded PLGA-HP-PEG-cRGD-platelet by mixing an exemplary formed PLGA-HP-PEG- cRGD-platelet with bFGF in a phosphate-buffered saline (PBS) solution, forming a stabilized bFGF loaded PLGA-HP-PEG-cRGD-platelet solution by adding at least one stabilizing agent to an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet solution, fluorescent labeling an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet by mixing at least one fluorescent label with bFGF loaded PLGA-HP-PEG-cRGD-platelet, and injecting an exemplary fluorescent labeled stabilized bFGF loaded PLGA-HP-PEG-cRGD-platelet solution into a PVA solution using a first syringe pump. In an exemplary embodiment, forming an exemplary core portion may include injecting liquid PFH into an exemplary PVA solution using a second syringe pump.
[0017] In an exemplary embodiment, injecting an exemplary predetermined amount of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles may include injecting an amount of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles in a range of 10 pg/mL to 2000 pg/mL intravenously to an exemplary body of an exemplary patient.
[0018] In an exemplary embodiment, forming PLGA-HP may include mixing dissolved heparin salt, NHS, and EDC with an exemplary formed PLGA-NH2 in DMF/FA at a temperature in a range of 4°C to 25°C for a time period in a range of 100 minutes to 2000 minutes. In an exemplary embodiment, injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution into an exemplary PVA solution may include injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution with a flow rate in a range of 100 pl/min to 500 pl/min into the PVA solution using an exemplary first syringe pump.
[0019] In an exemplary embodiment, injecting an exemplary liquid PFH into an exemplary PVA solution may include injecting an exemplary liquid PFH into an exemplary PVA solution with a flow rate in a range of 1 pl/min to 500 pl/min using an exemplary second syringe pump. In an exemplary embodiment, forming an exemplary PLGA-HP-PEG-cRGD-platelet may include mixing an exemplary platelet solution with an exemplary PLGA-HP-PEG-cRGD with a volume ratio in a range of 100:1 to 10:1 (PLGA-HP-PEG-cRGD: platelet). In an exemplary embodiment, forming an exemplary PLGA-HP-PEG-cRGD may include mixing an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA with an exemplary cRGD solution with a weight ratio in a range of 10000:1 to 100:1 (PLGA-HP-PEG-NHS: cRGD).
BRIEF DESCRIPTION OF THE DRAWINGS [0020] The drawing figures depict one or more implementations in accord with the present teachings, by way of example only, not by way of limitation. In the figures, like reference numerals refer to the same or similar elements.
[0021] FIG. 1A illustrates a flowchart of a method for healing myocardial infarction (MI), consistent with one or more exemplary embodiments of the present disclosure;
[0022] FIG. IB illustrates a flowchart of a method to produce an exemplary plurality of coreshell structured microbubbles, consistent with one or more exemplary embodiments of the present disclosure;
[0023] FIG. 2A illustrates a chemical reaction for synthesizing PLGA-NHS, consistent with one or more exemplary embodiments of the present disclosure;
[0024] FIG. 2B illustrates a chemical reaction for synthesizing PLGA-NH2, consistent with one or more exemplary embodiments of the present disclosure;
[0025] FIG. 2C illustrates a chemical reaction for synthesizing PLGA-HP, consistent with one or more exemplary embodiments of the present disclosure;
[0026] FIG. 2D illustrates a chemical reaction for synthesizing PLGA-HP-PEG-cRGD, consistent with one or more exemplary embodiments of the present disclosure;
[0027] FIG. 3 illustrates a microfluidic device for producing an exemplary plurality of coreshell structured microbubbles, consistent with one or more exemplary embodiments of the present disclosure;
[0028] FIG. 4 illustrates a spectrum of proton nuclear magnetic resonance (1HNMR) spectroscopy of PLGA-HP-PEG-NHS, consistent with one or more exemplary embodiments of the present disclosure;
[0029] FIG. 5 illustrates an optical microscopy image of microbubbles (MBs) obtained at flow rates of 1 pl/min for PFH and 250 pl/min for an exemplary bFGF loaded PLGA-HP-PEG- cRGD-platelet solution, consistent with one or more exemplary embodiments of the present disclosure;
[0030] FIG. 6A illustrates an image of MBs before applying LIFU, consistent with one or more exemplary embodiments of the present disclosure;
[0031] FIG. 6B illustrates an image of MBs after 20 minutes of applying LIFU with acoustic intensity of 35 mW/cm2, consistent with one or more exemplary embodiments of the present disclosure; [0032] FIG. 6C illustrates an image of MBs after 45 minutes of applying LIFU with acoustic intensity of 35 mW/cm2, consistent with one or more exemplary embodiments of the present disclosure;
[0033] FIG. 6D illustrates an image of MBs after 7 minutes of applying LIFU with acoustic intensity of 330 mW/cm2, consistent with one or more exemplary embodiments of the present disclosure;
[0034] FIG. 6E illustrates an image of MBs after 12 minutes of applying LIFU with acoustic intensity of 330 mW/cm2, consistent with one or more exemplary embodiments of the present disclosure; [0035] FIG. 7 illustrates fluorescence microscopic images of drug delivery samples for analyzing cellular uptake, consistent with one or more exemplary embodiments of the present disclosure; and
[0036] FIG. 8 illustrates fluorescence microscopic images of an exemplary plurality of fluorescent labeled core-shell structured microbubbles and an exemplary plurality of core-shell structured microbubbles with no label, consistent with one or more exemplary embodiments of the present disclosure.
DESCRIPTION OF EMBODIMENTS
[0037] In the following detailed description, numerous specific details are set forth by way of examples in order to provide a thorough understanding of the relevant teachings. However, it should be apparent that the present teachings may be practiced without such details. In other instances, well known methods, procedures, components, and/or circuitry have been described at a relatively high-level, without detail, in order to avoid unnecessarily obscuring aspects of the present teachings.
[0038] The novel features which are believed to be characteristic of the present disclosure, as to its structure, organization, use and method of operation, together with further objectives and advantages thereof, will be better understood from the following discussion. In the following detailed description, numerous specific details are set forth by way of examples in order to provide a thorough understanding of the relevant teachings. However, it should be apparent that the present teachings may be practiced without such details. In other instances, well known methods, procedures, components, and/or circuitry have been described at a relatively high- level, without detail, in order to avoid unnecessarily obscuring aspects of the present teachings. The following detailed description is presented to enable a person skilled in the art to make and use the methods and devices disclosed in exemplary embodiments of the present disclosure. For purposes of explanation, specific nomenclature is set forth to provide a thorough understanding of the present disclosure. However, it will be apparent to one skilled in the art that these specific details are not required to practice the disclosed exemplary embodiments. Descriptions of specific exemplary embodiments are provided only as representative examples. Various modifications to the exemplary implementations will be readily apparent to one skilled in the art, and the general principles defined herein may be applied to other implementations and applications without departing from the scope of the present disclosure. The present disclosure is not intended to be limited to the implementations shown, but is to be accorded the widest possible scope consistent with the principles and features disclosed herein.
[0039] The present disclosure is directed to exemplary embodiments of a drug delivery composition for healing myocardial infarction (MI) and a method to produce and administer an exemplary drug delivery composition. MI is a cardiovascular disease that is caused by a blockage in arteries that supply blood to heart. MI is a condition of heart failure due to insufficient blood (oxygen) supply to heart. In an exemplary embodiment, an exemplary drug delivery composition may carry a drug to a target place in body. In an exemplary embodiment, an exemplary target place may include platelet-clogged arteries that supply blood to heart. In an exemplary embodiment an exemplary drug delivery composition may be activated by an ultrasonic energy to release an exemplary drug. In an exemplary embodiment, an exemplary drug may include basic fibroblast growth factor (bFGF). In an exemplary embodiment, an exemplary ultrasonic energy may include a high-intensity focused ultrasound (HIFU) energy and a low-intensity focused ultrasound (LIFU) energy. In an exemplary embodiment, an exemplary LIFU energy may be used due to potential of an exemplary LIFU energy to focus on a specific site, minimize adverse side effects, and prevent damage to surrounding tissues of an exemplary target place. In an exemplary embodiment, exemplary adverse side effects may include producing undesired heating by HIFU energy. In an exemplary embodiment, an exemplary ultrasonic energy may be produced using an ultrasonic device applying an exemplary ultrasonic energy with acoustic frequency in a range of 1 MHz to 2 MHz. In an exemplary embodiment, an exemplary ultrasonic energy may be produced using an ultrasonic device applying an exemplary ultrasonic energy with acoustic intensity in a range of 1 mWcm"2 to 600 mWcm"2.
[0040] In an exemplary embodiment, an exemplary drug delivery composition may include a plurality of core-shell structured microbubbles. In an exemplary embodiment, an exemplary plurality of core-shell structured microbubbles may include a core portion and a shell portion. In an exemplary embodiment, an exemplary core portion may include liquid perfluorohexane (PFH). In an exemplary embodiment, liquid PFH may have a boiling point in a range of 58°C to 60°C. In an exemplary embodiment, an exemplary liquid PFH may be vaporized via an exemplary ultrasonic energy. In an exemplary embodiment, an exemplary ultrasonic energy may be produced utilizing an ultrasonic device. In an exemplary embodiment, an exemplary shell portion of each respective core-shell structured microbubble may include poly(D,L- lactide-co-glycolide)-heparin-polyethylene glycol-cyclic arginine glycine aspartate-platelet (PLGA-HP-PEG-cRGD-platelet), bFGF, at least a fluorescent label, and at least a stabilizing agent. In an exemplary embodiment, bFGF may be attached to an exemplary heparin of PLGA- HP-PEG-cRGD-platelet. In an exemplary embodiment, heparin may enhance bFGF loading on PLGA-HP-PEG-cRGD-platelet by interacting with bFGF. In an exemplary embodiment, heparin may have a negative electrical charge. In an exemplary embodiment, bFGF may have a positive electrical charge. In an exemplary embodiment, an ionic interaction may be formed between an exemplary negative electric charge of heparin and an exemplary positive electrical charge of bFGF. In an exemplary embodiment, RGD is a peptide that may be attached to GP Ilb/IIIa receptors. In an exemplary embodiment, GP Ilb/IIIa receptors may be formed on surface of activated platelets in blood vessels. In an exemplary embodiment, cyclic RGD (cRGD) may have greater affinity than linear RGD to GP Ilb/IIIa receptors. In an exemplary embodiment, an exemplary greater affinity of cRGD in comparison to linear RGD to GP Ilb/IIIa receptors may result in more stability of GP Ilb/IIIa receptor and cRGD. In an exemplary embodiment, platelet in PLGA-HP-PEG-cRGD-platelet may also increase accumulation of an exemplary plurality pf core-shell structured microbubbles at an exemplary target place. In an exemplary embodiment, platelet may have physical interaction with activated platelet in an exemplary target place. In an exemplary embodiment, PEG and PLGA may be biocompatible polymers for carrying bFGF, platelet, and cRGD. In an exemplary embodiment, dosage of bFGF may be monitored by controlling an amount of bFGF loaded on PLGA-HP- PEG-cRGD-platelet.
[0041] In an exemplary embodiment, an exemplary plurality of core- shell structured microbubbles may be dispersed in a polyvinyl alcohol (PVA) aqueous solution to form an emulsion. In an exemplary embodiment, an exemplary at least a stabilizing agent may be added to an exemplary shell portion for enhancing stability of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution. In an exemplary embodiment, an exemplary at least a stabilizing agent may include one of a plurality of gold nanoparticles (AuNPs), polyoxyethylene (20) sorbitan monooleate, and combinations thereof. In an exemplary embodiment, PVA may also enhance stability of an exemplary plurality of core- shell structured microbubbles in water. In an exemplary embodiment, an exemplary at least a stabilizing agent may include a plurality of AuNPs surrounded with chitosan (AuNPs- chitosan). In an exemplary embodiment, an exemplary chitosan layer may have a thickness in a range of 10 nm to 100 nm. In an exemplary embodiment, an exemplary at least a stabilizing agent may stabilize an exemplary plurality of core- shell structured microbubbles in an exemplary PVA solution. In an exemplary embodiment, an exemplary plurality of AuNPs- chitosan may stabilize an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution by inhibiting gas diffusion, and reducing interfacial tension of an exemplary shell, which may also limit Ostwald ripening. In an exemplary embodiment, as used herein “Ostwald ripening” may refer to growth of one emulsion droplet at expense of a smaller one as a result of difference in chemical potential of material within exemplary droplets. In an exemplary embodiment, polyoxyethylene (20) sorbitan monooleate may stabilize an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution by reducing sedimentation of an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution via enhancing viscosity of an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution.
[0042] In an exemplary embodiment, to track an exemplary drug delivery composition in body, an exemplary at least a fluorescent label may be added to PLGA-HP-PEG-cRGD-platelet. In an exemplary embodiment, an exemplary fluorescent label may include at least one of fluorescein isothiocyanate (FITC), rhodamine, and combinations thereof. In an exemplary embodiment, FITC may be added to PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 10:0.1 to 10:1 (PLGA-HP-PEG-cRGD-platelet: FITC). In an exemplary embodiment, FITC may have an excitation and an emission spectrum peak wavelengths of approximately 495 nm and 519 nm, respectively. In an exemplary embodiment, FITC may have a green color fluorescence emission.
[0043] In an exemplary embodiment, an exemplary drug delivery composition may include an emulsion of an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution. In an exemplary embodiment, an exemplary PVA solution may include an aqueous PVA solution with a concentration in a range of 0.1 w/w% to 5 w/w%. In an exemplary embodiment, an exemplary emulsion of an exemplary plurality of core- shell structured microbubbles in an exemplary PVA solution may include a weight ratio in a range of 10:1 to 2:1 (an exemplary plurality of core-shell structured microbubbles: PVA). In an exemplary embodiment, an exemplary plurality of core- shell structured microbubbles may be stable in an exemplary PVA solution for a time period in a range of 1 day to 60 days.
[0044] In an exemplary embodiment, a predetermined amount of an exemplary drug delivery composition may be injected intravenously to a patient suffering from MI. In an exemplary embodiment, an exemplary predetermined amount of an exemplary drug delivery composition may be in a range of 10 pL to 500 pL. In an exemplary embodiment, an exemplary drug delivery composition may reach an exemplary target place in an exemplary body of an exemplary patient. In an exemplary embodiment, cRGD may attach to GP Ub/IIIa receptors on activated platelet in arteries. In an exemplary embodiment, exemplary activated platelet may be accumulated inside arteries of an exemplary patient suffering from MI. In an exemplary embodiment, an exemplary target place may include platelet-clogged arteries that supply blood to heart. In an exemplary embodiment, an exemplary plurality of core-shell structured microbubbles may be systemically transferred using targeting ligands to an exemplary target place. In an exemplary embodiment, exemplary targeting ligands may include cRGD and platelet. In an exemplary embodiment, covalent or noncovalent attachment of exemplary targeting ligands on an exemplary shell may help recognizing antigens or receptors in an exemplary target place.
[0045] In an exemplary embodiment, after injecting an exemplary drug delivery composition intravenously to an exemplary patient, a fluorescent detector may be used to track FITC in an exemplary body of an exemplary patient. In an exemplary embodiment, an exemplary fluorescence detector may monitor fluorescent emission of an exemplary fluorescent label. In an exemplary embodiment, an exemplary fluorescent detector may be placed on chest of an exemplary patient. In an exemplary embodiment, an exemplary fluorescent detector may be placed on chest of an exemplary patent with a distance of an exemplary fluorescent detector to chest of an exemplary patient in a range of 1 cm to 5 cm h. In an exemplary embodiment, an exemplary fluorescent emission of an exemplary fluorescent label may be monitored utilizing an exemplary fluorescent detector for 20 hours to 72 hours. In an exemplary embodiment, tracking an exemplary fluorescent label may help to realize reaching an exemplary fluorescent label attached to PLGA-HP-PEG-cRGD-platelet to an exemplary target place.
[0046] In an exemplary embodiment, after tracking an exemplary drug delivery composition reaching an exemplary target place in an exemplary body of an exemplary patient, an exemplary LIFU energy may be applied utilizing an ultrasonic device to a chest of an exemplary patient. In an exemplary embodiment, an exemplary LIFU energy may be applied utilizing an ultrasonic device with a distance to a chest of a patient in a range of 1 cm to 5 cm. In an exemplary embodiment, an exemplary an exemplary LIFU energy may be applied on a chest of an exemplary patient for a time period in a range of 1 minute to 30 minutes. In an exemplary embodiment, an exemplary LIFU energy may include acoustic intensity in a range of 1 mWcm”2 to 600 mWcm”2. In an exemplary embodiment, an exemplary LIFU energy may include an acoustic frequency in a range of 1 MHz to 2 MHz. In an exemplary embodiment, an exemplary LIFU energy may induce an exemplary liquid PFH to evaporate. In an exemplary embodiment, evaporating an exemplary liquid PFH may increase internal pressure of an exemplary plurality of core-shell structured microbubbles. In an exemplary embodiment, an exemplary plurality of core-shell structures microbubbles may explode due to an exemplary increase of internal pressure of an exemplary plurality of core- shell structured microbubbles. Therefore, in an exemplary embodiment, bFGF may be released in an exemplary target place. In an exemplary embodiment, bFGF may induce blood vessel formation, promote proliferation restore perfusion and help migration of endothelial cells to infarcted area.
[0047] FIG. 1A illustrates a flowchart of a method 100 for healing myocardial infarction (MI), consistent with one or more exemplary embodiments of the present disclosure. In an exemplary embodiment, method 100 may include a step 102 of forming an emulsion of a plurality of coreshell structured microbubbles, step 104 of injecting a predetermined amount of the emulsion of the plurality of core-shell structured microbubbles intravenously to a body of a patient, step 106 of tracking a fluorescent label of each respective core- shell structured microbubble, and step 108 of applying an ultrasonic energy to chest of the patient.
[0048] In further detail with respect to step 102, step 102 of forming an emulsion of a plurality of core-shell structured microbubbles may include producing an exemplary plurality of coreshell structured microbubbles. FIG. IB illustrates a flowchart of a method 110 to produce an exemplary plurality of core-shell structured microbubbles, consistent with one or more exemplary embodiments of the present disclosure. In an exemplary embodiment, method 110 may include a step 102a of forming a shell portion of each core-shell structured microbubble of a plurality of core- shell structured microbubbles, and a step 102b of forming a core portion of each core- shell structured microbubble of the plurality of core- shell structured microbubbles.
[0049] In further detail with respect to step 102a, step 102a of forming an exemplary shell portion of each core-shell structured microbubble may include forming poly(D,L-lactide-co- glycolide)-NHS (PLGA-NHS) by dissolving PLGA-COOH, l-ethyl-3-(3- dimethylaminopropyl) carbodiimide (EDC), and NHS in an anhydrous dichloromethane solution. In an exemplary embodiment, PLGA-COOH may be mixed with EDC in a weight ratio in a range of 50: 1 to 10:1 (PLGA-COOH: EDC). In an exemplary embodiment, PLGA-COOH may be mixed with NHS in a weight ratio in a range of 50:1 to 5:1 (PLGA-COOH: NHS). In an exemplary embodiment, PLGA-COOH, EDC, and NHS may be mixed with an exemplary anhydrous dichloromethane solution with a weight ratio in a range of 1:1 to 1:2 (PLGA-COOH: an exemplary anhydrous dichloromethane solution). In an exemplary embodiment, PLGA-COOH, EDC, and NHS may be mixed with an exemplary anhydrous dichloromethane solution at a temperature in a range of 25°C to 35°C. In an exemplary embodiment, PLGA-COOH, EDC, and NHS may be mixed with an exemplary anhydrous dichloromethane solution under inert gas atmosphere. In an exemplary embodiment, an exemplary inert gas may include nitrogen. FIG. 2A illustrates a chemical reaction of synthesizing PLGA-NHS, consistent with one or more exemplary embodiments of the present disclosure.
[0050] In an exemplary embodiment, after forming PLGA-NHS, step 102a may further include forming PLGA-NH2 by dissolving formed PLGA-NHS and ethylenediamine (EDA) in a dichloromethane solution. In an exemplary embodiment, PLGA-NHS and EDA may be dissolved in an exemplary dichloromethane solution with a weight ratio in a range of 10:1 to 10:1.2 (PLGA-NHS: EDA). In an exemplary embodiment, PLGA-NHS and EDA may be dissolved in an exemplary dichloromethane solution at a temperature in a range of 25°C to 35°C. In an exemplary embodiment, PLGA-NHS and EDA may be mixed in an exemplary dichloromethane solution using a mixer with a rotational speed in a range of 100 rpm to 500 rpm. In an exemplary embodiment, PLGA-NHS and EDA may be mixed in an exemplary dichloromethane solution for 60 minutes to 2000 minutes. FIG 2B illustrates a chemical reaction of synthesizing PLGA-NH2, consistent with one or more exemplary embodiments of the present disclosure.
[0051] In further detail with respect to step 102a, after forming PLGA-NH2, step 102a may further include forming PLGA-HP by mixing dissolved heparin salt, NHS, and EDC in N,N- dimethylformamide/formamide (DMF/FA) with an exemplary formed PLGA-NH2. In an exemplary embodiment, a chemical reaction between carboxyl group of activated heparin and amine group of PLGA may happen. In an exemplary embodiment, an exemplary chemical reaction may lead to amide bond formation between PLGA and heparin. In an exemplary embodiment, DMF and FA may be mixed with a weight ratio in a range of 10:1 to 1:1 (DMF: FA). In an exemplary embodiment, heparin salt, NHS, EDC, and an exemplary formed PLGA-NH2 may be mixed in DMF/FA with a weight ratio in a range of 1:1 to 1:5 (heparin: NHS). In an exemplary embodiment, heparin salt, NHS, EDC, and an exemplary formed PLGA-NH2 may be mixed in DMF/FA with a weight ratio in a range of 10:1 to 1:1 (heparin: EDC). In an exemplary embodiment, heparin salt, NHS, EDC, and an exemplary formed PLGA-NH2 may be mixed in DMF/FA with a weight ratio in a range of 1:1 to 1:5 (heparin: an exemplary formed PLGA-NH2). In an exemplary embodiment, heparin salt, NHS, EDC, and an exemplary formed PLGA-NH2 may be mixed in DMF/FA at a temperature in a range of 25°C to 35°C. FIG 2C illustrates a chemical reaction of synthesizing PEGA-HP, consistent with one or more exemplary embodiments of the present disclosure
[0052] In further detail with respect to step 102a, step 102a may further include forming HOOC-PEG-COOH by dissolving HO-PEG-OH, succinic anhydride, and 4- (dimethylamino) pyridine (DMAP) in a dichloromethane solution. In an exemplary embodiment, HO-PEG-OH, succinic anhydride, and DMAP may be mixed in an exemplary dichloromethane solution utilizing a mixer with a rotational speed in a range of 100 rpm to 1000 rpm for time period in a range of 24 hours to 50 hours. In an exemplary embodiment, HO-PEG-OH, succinic anhydride, and DMAP may be mixed in an exemplary dichloromethane solution with a weight ratio in a range of 20:1 to 10:1 (HO-PEG-OH: DMAP). In an exemplary embodiment, HO-PEG-OH, succinic anhydride, and DMAP may be mixed in an exemplary dichloromethane solution with a weight ratio in a range of 2: 1 to 1 : 1 (HO-PEG-OH: succinic anhydride).
[0053] In further detail with respect to step 102a, after forming HOOC-PEG-COOH, step 102a may further include forming HOOC-PEG-NHS by dissolving an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an anhydrous dichloromethane solution. In an exemplary embodiment, dissolving an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an anhydrous dichloromethane solution may include mixing an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an exemplary anhydrous dichloromethane solution utilizing a mixer with a rotational speed in a range of 100 rpm to 1000 rpm. In an exemplary embodiment, dissolving an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an exemplary anhydrous dichloromethane solution may include mixing an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an exemplary anhydrous dichloromethane solution utilizing an exemplary mixer for a time period in a range of 15 hours to 50 hours. In an exemplary embodiment, dissolving an exemplary formed HOOC-PEG-COOH, EDC, and NHS in an anhydrous dichloromethane solution may be performed under an inert gas atmosphere to activate carboxylic groups of PEG. In an exemplary embedment, an exemplary inert gas may include nitrogen. [0054] In further detail with respect to step 102a, after forming HOOC-PEG-NHS, step 102a may further include forming PLGA-HP-PEG-NHS by mixing an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP and dicyclohexylcarbodiimide (DCC). In an exemplary embodiment, an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP may be mixed with dicyclohexylcarbodiimide (DCC) utilizing a mixer with a rotational speed in a range of 100 rpm to 1000 rpm. in an exempalry embodiment, an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP may be mixed with dicyclohexylcarbodiimide (DCC) utilizing an exempalry mixer for a time period in a range of 20 hours to 30 hours. In an exemplary embodiment, an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP may be mixed with DCC under an inert gas atmosphere. In an exemplary embodiment, an exemplary inert gas may include nitrogen. In an exemplary embodiment, an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP may be mixed with DCC with a weight ratio in a range of 1:1 to 1:10 (HOOC-PEG-NHS: PLGA-HP). In an exemplary embodiment, an exemplary formed HOOC-PEG-NHS and an exemplary formed PLGA-HP dissolved in DMF/FA with DMAP may be mixed with DCC with a weight ratio in a range of 100:1 to 10:1 (HOOC-PEG-NHS: DCC).
[0055] In further detail with respect to step 102a, after forming PLGA-HP-PEG-NHS, step 102a may further include forming PLGA-HP-PEG-cRGD by mixing an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA with a cyclic arginine-glycine-aspartate (cRGD) solution. In an exemplary embodiment, an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA may be added dropwise to an exemplary cRGD solution. In an exemplary embodiment, an exemplary cRGD solution may have a concentration in a range of 1 mol/L to 100 mol/L. In an exemplary embodiment, an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA may be mixed with an exemplary cRGD solution utilizing a mixer with a rotational speed in a range of 1 rpm to 10 rpm. In an exemplary embodiment, an exemplary formed PLGA-HP-PEG-NHS solution in DMF/FA may be mixed with an exemplary cRGD solution utilizing an exemplary mixer for a time period in a range of 200 minutes to 2000 minutes. FIG 2D illustrates a chemical reaction for synthesizing PLGA-HP-PEG-cRGD, consistent with one or more exemplary embodiments of the present disclosure. [0056] In further detail with respect to step 102a, after forming PLGA-HP-PEG-cRGD, step 102a may further include forming PLGA-HP-PEG-cRGD-platelet by mixing a platelet solution with an exemplary formed PLGA-HP-PEG-cRGD. In an exemplary embodiment, platelet may be separated from blood by differential centrifugation. In an exemplary embodiment, blood may be centrifuged with a rotational speed in a range of 2000 rpm to 4000 rpm for a time period in a range of 10 minutes to 30 minutes. In an exemplary embodiment, an exemplary platelet solution may be mixed with an exemplary formed PLGA-HP-PEG-cRGD utilizing a mixer with a rotational speed in a range of 100 rpm to 500 rpm. In an exemplary embodiment, an exemplary platelet solution may be mixed with an exemplary formed PLGA-HP-PEG-cRGD utilizing an exemplary mixer for a time period in a range of 60 minutes to 2000 minutes. In an exemplary embodiment, an exemplary platelet solution may be mixed with an exemplary formed PLGA-HP-PEG-cRGD with a weight ratio in a range of 100:1 to 10:1 (PLGA-HP- PEG-cRGD: platelet).
[0057] In further detail with respect to step 102a, after forming PLGA-HP-PEG-cRGD- platelet, step 102a may further include forming bFGF loaded PLGA-HP-PEG-cRGD-platelet by mixing an exemplary formed PLGA-HP-PEG-cRGD-platelet with bFGF dissolved in a phosphate-buffered saline (PBS) solution. In an exemplary embodiment, an exemplary formed PLGA-HP-PEG-cRGD-platelet may be mixed with an exemplary bFGF solution utilizing a mixer with a rotational speed in a range of 100 rpm to 500 rpm. In an exemplary embodiment, an exemplary formed PLGA-HP-PEG-cRGD-platelet may be mixed with an exemplary bFGF solution for a time period in a range of 100 minutes to 3000 minutes. In an exemplary embodiment, an exemplary PBS solution may have a concentration in a range of 1 mol/L to 10 mol/L. In an exemplary embodiment, an exemplary bFGF solution may have a concentration in a range of 1 mol/L to 100 mol/L. In an exemplary embodiment, an exemplary formed PLGA- HP-PEG-cRGD-platelet may be mixed with an exemplary bFGF solution with a weight ratio in a range of 10:1 to 2:1 (an exemplary formed PLGA-HP-PEG-cRGD-platelet: bFGF).
[0058] In further detail with respect to step 102a, after forming an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet, step 102a may further include forming a plurality of fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet structures. In an exemplary embodiment, forming an exemplary plurality of fluorescent labeled bFGF loaded PLGA-HP- PEG-cRGD-platelet structures may include mixing at least a fluorescent label with bFGF loaded PLGA-HP-PEG-cRGD-platelet. In an exemplary embodiment, an exemplary at least a fluorescent label may include at least one of fluorescein isothiocyanate (FITC), rhodamine, and combinations thereof. In an exemplary embodiment, FITC may be attached to bFGF. In an exemplary embodiment, FITC may be mixed with an exemplary bFGF loaded PLGA-HP-PEG- cRGD-platelet structures with a weight ratio in a range of 1:10 to 1:100 (FITC: an exemplary plurality of bFGF loaded PLGA-HP-PEG-cRGD-platelet structures).
[0059] In further detail with respect to step 102a, step 102a may further include adding at least a stabilizing agent to bFGF loaded PLGA-HP-PEG-cRGD-platelet. In an exemplary embodiment, an exemplary at least a stabilizing agent may be mixed with bFGF loaded PLGA- HP-PEG-cRGD-platelet utilizing a mixer with a rotational speed in a range of 100 rpm to 1000 rpm. In an exemplary embodiment, an exemplary at least a stabilizing agent may be mixed with bFGF loaded PLGA-HP-PEG-cRGD-platelet utilizing an exemplary mixer for a time period in a range of 1200 minutes to 2500 minutes. In an exemplary embodiment, an exemplary stabilizing agent may include at least one of a plurality of gold nanoparticles (AuNPs), polyoxyethylene (20) sorbitan monooleate, and combinations thereof. In an exemplary embodiment, an exemplary plurality of AuNPs may be synthesized using chitosan as a reducing agent. In an exemplary embodiment, an exemplary chitosan may be wrapped over an exemplary plurality of AuNPs (AuNPs-chitosan). In an exemplary embodiment, an exemplary plurality of AuNPs may be synthesized by heating an aqueous solution of HAuCL and a chitosan solution at a temperature in a range of 40°C to 100°C for a time period in a range of 30 minutes to 300 minutes. In an exemplary embodiment, each respective AuNP-chitosan may include an AuNP surrounded with a layer of chitosan with a thickness in a range of 10 nm to 100 nm. In an exemplary embodiment, an exemplary plurality of core- shell structured microbubbles may include polyoxyethylene (20) sorbitan monooleate with a weight ratio in a range of 1:10 to 1:1 (polyoxyethylene (20) sorbitan monooleate: PLGA-HP-PEG-cRGD- platelet). In an exemplary embodiment, an exemplary plurality of AuNPs may be attached to an exemplary shell of an exemplary plurality of core-shell structured microbubbles. In an exemplary embodiment, an exemplary plurality of core- shell structured microbubbles may include an exemplary plurality of AuNPs-chitosan with a weight ratio in a range of 1:1000 to 1:10 (Au NPs-chitosan: PLGA-HP-PEG-cRGD-platelet). In an exemplary embodiment, an exemplary plurality of core- shell structured microbubbles may include polyoxyethylene (20) sorbitan monooleate with a weight ratio in a range of 1 : 10 to 1:1 (polyoxyethylene (20) sorbitan monooleate: PLGA-HP-PEG-cRGD-platelet). [0060] In an exemplary embodiment, step 102a may further include fluorescent labeling an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet with at least a fluorescent label. In an exemplary embodiment, an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet may be fluorescent labeled by mixing an exemplary at least a fluorescent label with bFGF loaded PLGA-HP-PEG-cRGD-platelet. In an exemplary embodiment, an exemplary fluorescent label may include at least one of fluorescein isothiocyanate (FITC), rhodamine, and combinations thereof. In an exemplary embodiment, FITC may attach to bFGF. In an exemplary embodiment, a FITC solution in dimethyl sulfoxide (DMSO) may be added to an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:20 to 1:10 (FITC: bFGF loaded PLGA-HP-PEG-cRGD-platelet). In an exemplary embodiment, an exemplary FITC solution and an exemplary bFGF loaded PLGA-HP-PEG-cRGD-platelet may be mixed for a time period in a range of 60 minutes to 1800 minutes. In an exemplary embodiment, an exemplary FITC solution and an exemplary bFGF loaded PLGA-HP-PEG- cRGD-platelet may be mixed in darkness.
[0061] In further detail with respect to step 102a, step 102a may further include injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution into a PVA solution using a first syringe pump. In an exemplary embodiment, an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution may be injected into a microfluidic device. In an exemplary embodiment, an exemplary PVA solution may be injected into an exemplary microfluidic device before injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution. In an exemplary embodiment, injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution may include injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG- cRGD-platelet solution into an exemplary PVA solution using an exemplary first syringe pump with a flow rate in a range of 100 pl/min to 500 pl/min. In an exemplary embodiment, an exemplary PVA solution may have a concentration in a range of 0.1 w/w% to 5 w/w% in water. In an exemplary embodiment, an exemplary solution of bFGF loaded PLGA-HP-PEG-cRGD- platelet in phosphate-buffered saline (PBS) may be injected into a microfluidic device utilizing an exemplary first syringe pump.
[0062] In further detail with respect to step 102b, step 102b of forming a core portion of each core- shell structured microbubble may include injecting a liquid perfluorohexane (PFH) into an exemplary PVA solution. In an exemplary embodiment, an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution and an exemplary liquid PFH may be injected into an exemplary PVA solution, simultaneously. In an exemplary embodiment, an exemplary liquid PFH may be injected into an exemplary microfluidic device utilizing a second syringe pump. In an exemplary embodiment, an exemplary liquid PFH may be injected into an exemplary PVA solution with a flow rate in a range of 1 pl/min to 500 pl/min. In an exemplary embodiment, an exemplary fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution and an exemplary liquid PFH may be injected into an exemplary PVA solution with a weight ratio in a range of 1:10 to 1:1 (bFGF loaded PLGA-HP-PEG-cRGD-platelet solution: liquid PFH). In an exemplary embodiment, an exemplary PVA solution may have a concentration in a range of 0.1 w/w% to 5 w/w% in water.
[0063] Referring back to FIG. 1A, in further detail with respect to step 102, step 102 of forming an emulsion of a plurality of core- shell structured microbubbles may include stabilizing an exemplary emulsion of a plurality of core- shell structured microbubbles in a PVA solution. In an exemplary embodiment, each respective core-shell structured microbubble of an exemplary plurality of core-shell structured microbubbles may have a size in a range of 1 pm to 30 pm. In an exemplary embodiment, an exemplary emulsion of a plurality of core-shell structured microbubbles may be stabilized sing at least a stabilizing agent. In an exemplary embodiment, an exemplary at least a stabilizing agent may include at least one of a plurality of gold nanoparticles (AuNPs), polyoxyethylene (20) sorbitan monooleate, and combinations thereof. In an exemplary embodiment, polyoxyethylene (20) sorbitan monooleate may be attached to an exemplary shell of an exemplary plurality of core- shell structured microbubbles with a weight ratio in a range of 1:10 to 1:1 (polyoxyethylene (20) sorbitan monooleate: an exemplary shell). In an exemplary embodiment, polyoxyethylene (20) sorbitan monooleate may be physically attached on an exemplary shell. In an exemplary embodiment, an exemplary plurality of AuNPs may be attached to an exemplary shell of an exemplary plurality of coreshell structured microbubbles. In an exemplary embodiment, an exemplary plurality of AuNPs may stabilize an exemplary emulsion by inhibition of gas diffusion to an exemplary plurality of core- shell structured microbubbles due to presence of solid AuNPs. In an exemplary embodiment, an exemplary plurality of AuNPs may stabilize an exemplary emulsion by reducing interfacial tension which may also limit Ostwald ripening. In an exemplary embodiment, an exemplary plurality of AuNPs may be produced using chitosan as a reducing agent. In an exemplary embodiment, each respective AuNP may include a layer of chitosan wrapped over an exemplary AuNP (AuNP-chitosan). In an exemplary embodiment, an exemplary layer of chitosan may have a thickness in a range of 10 nm to 100 nm. In an exemplary embodiment, AuNPs- chitosan may be produced by mixing an aqueous solution of HAuCh with an aqueous solution of chitosan in a mixer with a rotational speed in a range of 100 rpm to 1000 rpm. In an exemplary embodiment, an exemplary solution of HAuCU may be added to an exemplary aqueous solution of chitosan with a weight ratio in a range of 1:100 to 1:10 (chitosan: HAuCU). In an exemplary embodiment, exemplary AuNPs-chitosan may be produced at a temperature in a range of 30°C to 100°C. In an exemplary embodiment, an exemplary plurality of AuNPs-chitosan may be attached to PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:1000 to 1:10 (Au NPs-chitosan: PLGA-HP-PEG-cRGD- platelet). In an exemplary embodiment, each respective AuNP of an exemplary plurality of AuNPs may have a diameter in a range of 25 nm to 75 nm. In an exemplary embodiment, PVA may also stabilize an exemplary emulsion by reducing sedimentation of an exemplary plurality of core-shell structured microbubbles via enhancing viscosity of an exemplary emulsion.
[0064] FIG. 3 illustrates a microfluidic device 300 for producing an exemplary plurality of core- shell structured microbubbles, consistent with one or more exemplary embodiments of the present disclosure. In an exemplary embodiment, microfluidic device 300 may include a plate 312, a microfluidic vessel 310, a first inlet 302, a second inlet 304, a third inlet 306, and an outlet 308. In an exemplary embodiment, plate 312 may be made of at least one of polydimethyl siloxane (PDMS), glass, or polymethyl methacrylate (PMMA). In an exemplary embodiment, plate 312 may be a platform for holding microfluidic vessel 310, first inlet 302, second inlet 304, third inlet 306, and outlet 308 thereon. In an exemplary embodiment, first inlet 302 may be used for injecting an exemplary fluorescent labeled bFGF loaded PLGA-HP- PEG-cRGD-platelet solution into microfluidic vessel 310 utilizing an exemplary first syringe pump. In an exemplary embodiment, second inlet 304 may be used for injecting an exemplary liquid PFH into microfluidic vessel 310 utilizing an exemplary second syringe pump. In an exemplary embodiment, an exemplary PVA solution may be injected via third inlet 306 into microfluidic vessel 310. In an exemplary embodiment, outlet 308 may be used to collect an exemplary drug delivery composition produced by method 100 and method 110. In an exemplary embodiment, an exemplary drug delivery composition may be an exemplary emulsion of an exemplary plurality of core-shell structured microbubbles in an exemplary PVA solution. In an exemplary embodiment, an exemplary PVA solution may have a concentration in a range of 0.1 to 5 wlwc/c in water. In an exemplary embodiment, each respective core- shell structured microbubble of an exemplary plurality of core- shell structured microbubbles may have a size in a range of 1 pm to 100 pm.
[0065] Referring back to FIG. 1A, in further detail with respect to step 104, step 104 of injecting a predetermined amount of an exemplary drug delivery composition may include injecting an exemplary predetermined amount of an exemplary drug delivery composition intravenously to a body of a patient utilizing an ampule. In an exemplary embodiment, injecting an exemplary predetermined amount of an exemplary drug delivery composition may include injecting an amount of an exemplary drug delivery composition in a range of 10 pL to 500 pL intravenously to an exemplary body of an exemplary patient. In an exemplary embodiment, an exemplary drug delivery composition may be injected intravenously to a body of a patient after a time period in a range of 60 minutes to 7200 minutes after producing an exemplary drug delivery composition due to coagulation of an exemplary plurality of core-shell structured microbubbles. In an exemplary embodiment, an exemplary drug delivery composition may be injected intravenously to a body of a human and a body of an animal.
[0066] In further detail with respect to step 106, step 106 of tracking a fluorescent label of each respective core- shell structured microbubble may include monitoring an exemplary fluorescent label of each respective core-shell structured microbubble. In an exemplary embodiment, tracking an exemplary fluorescent label of each respective core-shell structured microbubble may include tracking an exemplary fluorescent label of each respective core-shell structured microbubble utilizing a fluorescent detector. In an exemplary embodiment, an exemplary drug delivery composition may include at least a fluorescent label. In an exemplary embodiment, an exemplary fluorescent label may include at least one of FITC, rhodamine, and combinations thereof. In an exemplary embodiment, an exemplary fluorescent label attached to bFGF in an exemplary drug delivery composition may be monitored utilizing an exemplary fluorescent detector after injecting an exemplary drug delivery composition intravenously to a body of an exemplary patient. In an exemplary embodiment, bFGF may be attached to FITC for monitoring in vivo migration of an exemplary plurality of core- shell structured microbubbles through blood vessels. In an exemplary embodiment, FITC may emit light with a wavelength in a green region. In an exemplary embodiment, an exemplary fluorescent detector may show existence of an exemplary core-shell structured microbubbles using fluorescence emission of FITC. In an exemplary embodiment, an exemplary fluorescence detector may be placed on chest of an exemplary patient. In an exemplary embodiment, accumulation of FITC in an exemplary target place may be understood when an exemplary target place turns to a green color (shown by an exemplary fluorescent detector). In an exemplary embodiment, FITC may be attached to bFGF of an exemplary drug delivery composition. Therefore, green color emission in an exemplary target place may show existence of an exemplary drug delivery composition in an exemplary target place.
[0067] In an exemplary embodiment, after accumulation of FITC in an exemplary target place identified by an exemplary fluorescent detector, an ultrasonic device may be placed on chest of an exemplary patient. In further detail with respect to step 108, step 108 of applying an ultrasonic energy to chest of an exemplary patient may include applying an exemplary ultrasonic energy in a distance in a range of 1 cm to 5 cm to chest of an exemplary patient. In an exemplary embodiment, after reaching an exemplary drug delivery composition to an exemplary target place, an exemplary ultrasonic energy may be applied on chest of an exemplary patient. In an exemplary embodiment, an exemplary drug delivery composition may reach to an exemplary target place in a time period of maximum 72 hours. In an exemplary embodiment, an exemplary ultrasonic energy may be an acoustic energy with acoustic frequency in a range of 1 MHz to 2 MHz. In an exemplary embodiment, an exemplary ultrasonic energy may be applied utilizing an ultrasonic device with acoustic intensity in a range of 1 mWcm”2 to 600 mWcm"2. In an exemplary embodiment, applying an exemplary ultrasonic energy may include applying an exemplary ultrasonic energy on an exemplary chest of an exemplary patient for a time period in a range of 1 minute to 30 minutes. In an exemplary embodiment, applying an exemplary ultrasonic energy to chest of an exemplary patient may include applying an exemplary ultrasonic energy to chest of an exemplary patient after reaching an exemplary drug delivery composition to an exemplary target place. In an exemplary embodiment, an exemplary target place may include platelet-clogged arteries that supply blood to heart.
[0068] In an exemplary embodiment, after applying an exemplary ultrasonic device to chest of an exemplary patient, an exemplary liquid PFH may vaporize. In an exemplary embodiment, an exemplary plurality of core-shell structured micro bubbles may get larger with an exemplary vaporization process. In an exemplary embodiment, an exemplary plurality of core-shell structured micro bubbles may expand until an exemplary core-shell structured micro bubbles may lose an exemplary core-shell structure and may explode. In an exemplary embodiment, an exemplary plurality of core-shell structured micro bubbles may lose an exemplary core-shell structure after a time period in a range of 1 minute to 120 minutes of ultrasonic illumination. In an exemplary embodiment, after exploding an exemplary plurality of core-shell structured microbubbles, bFGF may be released in an exemplary target place. In an exemplary embodiment, an exemplary slow process of explosion may help to controlled release of bFGF. In an exemplary embodiment, after a time period in a range of 1 minute to 5000 minutes, all of an exemplary plurality of core-shell structured micro bubbles may explode. In an exemplary embodiment, bFGF may help blood vessel formation, promote proliferation restore perfusion and help migration of endothelial cells in infarcted area.
[0069] Example 1: Producing PLGA-HP-PEG-cRGD-platelet
[0070] For producing PLGA-HP, a method similar to step 102 of method 100 and/or method 110 may be used. To produce PLGA-HP, low molecular weight heparin salt (0.02 mmol), NHS (0.27 mmol), and EDC (0.05 mmol) were dissolved in DMF/FA (5 mL). In the next step, PLGA-NH2 (0.004 mmol) was reacted at room temperature. Then, the product was dialyzed against distilled water for three days. The product was then lyophilized to obtain PLGA-HP. PLGA-HP conjugated HOOC-PEG-NHS was obtained by a direct coupling reaction. Then, PLGA-HP (0.005 mmol) and HOOC-PEG-NHS (0.01 mmol) were dissolved in the mixture of DMF/FA solvent. Then, DMAP and DCC were added to the solution. The mixture of PLGA-HP, HOOC-PEG-NHS, DMAP and DCC (reaction mixture) were mixed.
[0071] The reaction mixture was precipitated in cold diethyl ether after the mixing process. A solution of PLGA-HP-PEG-NHS in anhydrous DMF/FA was poured dropwise into the cRGD solution. After stirring, the obtained solution was dialyzed to remove the unreacted mixture. The conjugate PLGA-HP-PEG-cRGD was then lyophilized. Finally, platelets were separated from whole sheep blood by differential centrifugation. The plasma blood bag was poured into a falcon and centrifuged to remove RBC. Then, platelet-rich plasma (PRP) was prepared by adding blood plasma into a tube and centrifuging the blood plasma. Platelets were re-suspended in PBS using anticoagulant ACD. Then, the mixture of platelets and PBS were added to PLGA-HP-PEG-cRGD.
[0072] Example 2: Producing microbubbles of bFGF-loaded PLGA-HP-PEG-cRGD- [0073] For producing droplets of bFGF-loaded PLGA-HP-PEG-cRGD-platelet, a method similar to method 100 and method 110 was used. For the first step, PLGA-HP-PEG-cRGD (50 mg) and bFGF (50 pg) were suspended in PBS. Then, bFGF-loaded PLGA-HP-PEG-cRGD was collected by centrifugation and washed with PBS to remove free bFGF. Then, bFGF- loaded PLGA-HP-PEG-cRGD (300 pL) and gold nanoparticles (300 pL) were ultrasonicated, and polyoxyethylene (20) sorbitan monooleate was added to the mixture. The obtained solution and perfluorohexane were injected using two independent syringe pumps at a predetermined constant flow rate. The obtained droplets from the microfluidic device were stabilized in a PVA solution and collected.
[0074] Microfluidic devices allow controlling the flow rates of two different solutions by employing two independent pumps. The inner solution was a perfluorohexane (PFH). PFH was used to prevent the coalescence of the particles and enhance particle stability. The outer aqueous phase was the polymeric solution containing PLGA-HP-PEG-cRGD/gold nanoparticles/ polyoxyethylene (20) sorbitan monooleate mixture. Droplets were made by using a microfluidic device demonstrated in FIG. 3. The final droplets were collected in a PVA solution as a stabilizing agent.
[0075] FIG. 4 illustrates a spectrum of proton nuclear magnetic resonance (1HNMR) spectroscopy of PLGA-HP-PEG-NHS, consistent with one or more exemplary embodiments of the present disclosure. The 1H-NMR spectrum of the final product displayed successful synthesis of PLGA-HP-PEG copolymer in which the main signals were related to PLGA moiety (a), (b), (c) at 1.46, 4.8, and 5.2 ppm of the lactide units in PLGA, respectively. The chemical shifts at 3.38 and 4.1 ppm (d, e) were related to the NH-CH2-CH2-NH, respectively. Also, the chemical shifts at 2.1 ppm (f), 3.1, and 3.5 ppm (g, h) were indicatives of CH-NH of heparin and PEG moiety (methylene hydrogen of the PEG), respectively (FIG. 4).
[0076] FIG. 5 illustrates an optical microscopy image of microbubbles (MBs) obtained at flow rates of 1 pl/min for PFH and 250 pl/min for an exemplary bFGF loaded PLGA-HP-PEG- cRGD-platelet solution, consistent with one or more exemplary embodiments of the present disclosure. PFH-droplets are interesting for ultrasound-triggered delivery systems. To disrupt the PFH-loaded microspheres were exposed to low intensity focused ultrasound (LIFU) (an acoustic frequency of 1.5 MHz and an acoustic intensity of 35 mW/cm2). When exposing the droplets to LIFU, the variations of the formed micro bubbles (MBs) were observed via light microscopy. The volume of the MBs increased by acoustic intensity of 330 mW/cm2, which conferred to an increasing number of droplets vaporized, and more MBs formed at 12 minutes. FIG. 6A illustrates an image of MBs before applying LIFU, consistent with one or more exemplary embodiments of the present disclosure. FIG. 6B illustrates an image of MBs after 20 minutes of applying LIFU with acoustic intensity of 35 mW/cm2, consistent with one or more exemplary embodiments of the present disclosure. FIG. 6C illustrates an image of MBs after 45 minutes of applying LIFU with acoustic intensity of 35 mW/cm2, consistent with one or more exemplary embodiments of the present disclosure. FIG. 6D illustrates an image of MBs after 7 minutes of applying LIFU with acoustic intensity of 330 mW/cm2, consistent with one or more exemplary embodiments of the present disclosure. FIG. 6E illustrates an image of MBs after 12 minutes of applying LIFU with acoustic intensity of 330 mW/cm2, consistent with one or more exemplary embodiments of the present disclosure.
[0077] The targeting specificity of bFGF-platelet-droplet for platelet-receptors was visually compared with non-targeted by fluorescence microscopic. Strong green fluorescence can be detected in the cells incubated with the targeted droplet. FIG. 7 illustrates fluorescence microscopic images of drug delivery samples for analyzing cellular uptake, consistent with one or more exemplary embodiments of the present disclosure. In FIG. 7 UTS stands for ultrasound irradiation. As shown in FIG. 7, different samples were tested with UTS and without UTS. Results showed that control sample (702), bFGF sample (704), bFGF- PLGA-HP-PEG-cRGD sample (706), and bFGF- PLGA-HP-PEG-cRGD-platelet sample (708) were illuminated by an ultrasonic energy. Control sample (710), bFGF sample (712), bFGF- PLGA-HP-PEG-cRGD sample (714), and bFGF- PLGA-HP-PEG-cRGD-platelet sample (716) were analyzed before an ultrasonic illumination. Results showed cellular uptake of bFGF- PLGA-HP-PEG-cRGD- platelet samples was higher due to interaction between platelet and activated platelet in blood vessels. These results confirmed the poor permeability of bFGF without MB as the carrier
[0078] Example 3: Analyzing in-vivo targeting ability of an exemplary drug delivery
Figure imgf000028_0001
[0079] To analyze in-vivo targeting ability of an exemplary drug delivery composition a fluorescent detector was used. In-vivo targeting images of FITC-bFGF-loaded PLGA-HP- PEG-cRGD- droplet and FITC-bFGF-loaded PLGA-HP-PEG-cRGD-droplet were accomplished using the fluorescence assessments. To study the targeting efficiency of the droplets, two model rats were randomly selected and shaved. Myocardial ischemia was induced in one model of rats. Then, FITC-bFGF-loaded PLGA HP-PEG-cRGD-platelet-droplet and FITC-bFGF-loaded PLGA-HP-PEG-cRGD-droplet (100 pL) were injected via the tail vein immediately. In-vivo fluorescence imaging of the heart was acquired post-injection and the relative fluorescence intensities were recorded. FIG. 8 illustrates fluorescence microscopic images of a plurality of fluorescent labeled core- shell structured microbubbles (804) and a plurality of core-shell structured microbubbles with no label (802), consistent with one or more exemplary embodiments of the present disclosure. As shown in FIG. 8, the plurality of fluorescent labeled core- shell structured microbubbles had fluorescent images with green color indicating the presence of the plurality of core-shell structured microbubbles.
[0080] Industrial Applicability
[0081] The present disclosure may include a drug delivery composition for healing MI. An exemplary drug delivery composition may include an emulsion of a plurality core- shell structured microbubble in a PVA solution. An exemplary emulsion may be injected intravenously to a patient. An exemplary drug delivery composition may be activated by an ultrasonic energy applied to chest of an exemplary patient. An exemplary drug delivery composition may be used in hospitals and clinics to treat MI and hinder side effects of oxygen deficiency of heart.
[0082] While the foregoing has described what are considered to be the best mode and/or other examples, it is understood that various modifications may be made therein and that the subject matter disclosed herein may be implemented in various forms and examples, and that the teachings may be applied in numerous applications, only some of which have been described herein. It is intended by the following claims to claim any and all applications, modifications and variations that fall within the true scope of the present teachings.
[0083] Unless otherwise stated, all measurements, values, ratings, positions, magnitudes, sizes, and other specifications that are set forth in this specification, including in the claims that follow, are approximate, not exact. They are intended to have a reasonable range that is consistent with the functions to which they relate and with what is customary in the art to which they pertain.
[0084] The scope of protection is limited solely by the claims that now follow. That scope is intended and should be interpreted to be as broad as is consistent with the ordinary meaning of the language that is used in the claims when interpreted in light of this specification and the prosecution history that follows and to encompass all structural and functional equivalents. Notwithstanding, none of the claims are intended to embrace subject matter that fails to satisfy the requirement of Sections 101, 102, or 103 of the Patent Act, nor should they be interpreted in such a way. Any unintended embracement of such subject matter is hereby disclaimed.
[0085] Except as stated immediately above, nothing that has been stated or illustrated is intended or should be interpreted to cause a dedication of any component, step, feature, object, benefit, advantage, or equivalent to the public, regardless of whether it is or is not recited in the claims.
[0086] It will be understood that the terms and expressions used herein have the ordinary meaning as is accorded to such terms and expressions with respect to their corresponding respective areas of inquiry and study except where specific meanings have otherwise been set forth herein. Relational terms such as first and second and the like may be used solely to distinguish one entity or action from another without necessarily requiring or implying any actual such relationship or order between such entities or actions. The terms “comprises,” “comprising,” or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. An element proceeded by “a” or “an” does not, without further constraints, preclude the existence of additional identical elements in the process, method, article, or apparatus that comprises the element.
[0087] The Abstract of the Disclosure is provided to allow the reader to quickly ascertain the nature of the technical disclosure. It is submitted with the understanding that it will not be used to interpret or limit the scope or meaning of the claims. In addition, in the foregoing Detailed Description, it can be seen that various features are grouped together in various implementations. This is for purposes of streamlining the disclosure, and is not to be interpreted as reflecting an intention that the claimed implementations require more features than are expressly recited in each claim. Rather, as the following claims reflect, inventive subject matter lies in less than all features of a single disclosed implementation. Thus, the following claims are hereby incorporated into the Detailed Description, with each claim standing on its own as a separately claimed subject matter.
[0088] While various implementations have been described, the description is intended to be exemplary, rather than limiting and it will be apparent to those of ordinary skill in the art that many more implementations and implementations are possible that are within the scope of the implementations. Although many possible combinations of features are shown in the accompanying figures and discussed in this detailed description, many other combinations of the disclosed features are possible. Any feature of any implementation may be used in combination with or substituted for any other feature or element in any other implementation unless specifically restricted. Therefore, it will be understood that any of the features shown and/or discussed in the present disclosure may be implemented together in any suitable combination. Accordingly, the implementations are not to be restricted except in light of the attached claims and their equivalents. Also, various modifications and changes may be made within the scope of the attached claims.

Claims

What is claimed is:
1. A drug delivery composition for healing myocardial infarction (MI), the drug delivery composition comprising: a plurality of core-shell structured microbubbles, each respective core-shell structured microbubble comprising: a core, comprising liquid perfluorohexane (PFH); and a shell surrounding the core, the shell comprising: poly(D,L-lactide-co-glycolide)-heparin-polyethylene glycolcyclic arginine glycine aspartate-platelet (PLGA-HP-PEG-cRGD- platelet); basic fibroblast growth factor (bFGF) attached to heparin of the PLGA-HP-PEG-cRGD-platelet with a weight ratio in a range of 1:10000 to 1:10 (bFGF: PLGA-HP-PEG-cRGD-platelet), wherein the liquid PFH has a weight ratio to the PLGA-HP-PEG- cRGD-platelet in a range of 1:10 to 1:1 (liquid PFH: PLGA-HP-PEG- cRGD-platelet); and at least one stabilizing agent attached to PLGA-HP-PEG-cRGD- platelet; and at least one fluorescent label attached to bFGF; and a polyvinyl alcohol (PVA) solution, the plurality of core- shell structured microbubbles being dispersed in the PVA solution.
2. The drug delivery composition of claim 1, wherein the at least one stabilizing agent comprises at least one of a plurality of gold nanoparticles (AuNPs), polyoxyethylene (20) sorbitan monooleate, and combination;
3. The drug delivery composition of claim 2, wherein each respective AuNP of the plurality of AuNPs has a particle size in a range of 25 nm to 75 nm.
4. The drug delivery composition of claim 2, wherein the at least one stabilizing agent comprises a plurality of AuNPs surrounded by chitosan (AuNPs-chitosan), each respective AuNP-chitosan comprising an AuNP surrounded with a layer of chitosan with a thickness in a range of 10 nm to 100 nm.
5. The drug delivery composition of claim 4, wherein the drug delivery composition comprises the plurality of AuNPs-chitosan with a weight ratio to the PLGA-HP-PEG-cRGD- platelet in a range of 1:1000 to 1:10 (Au NPs-chitosan: PLGA-HP-PEG-cRGD-platelet).
6. The drug delivery composition of claim 2, wherein the drug delivery composition comprises polyoxyethylene (20) sorbitan monooleate with a weight ratio to the PLGA-HP- PEG-cRGD-platelet in a range of 1:10 to 1:1 (polyoxyethylene (20) sorbitan monooleate: PLGA-HP-PEG-cRGD-platelet) .
7. The drug delivery composition of claim 1, wherein the PVA solution comprises an aqueous PVA solution with a concentration of PVA in a range of 0.1 w/w% to 5 w/w%.
8. The drug delivery composition of claim 1, wherein the at least one fluorescent label comprises at least one of fluorescein isothiocyanate, rhodamine, and combinations thereof.
9. The drug delivery composition of claim 1 , wherein each respective core-shell structured microbubble of the plurality of core- shell structured microbubbles has a size in a range of 1 pm to 30 pm.
10. A method for treating myocardial infarction (MI), the method comprising: forming an emulsion of a plurality of core- shell structured microbubbles, each respective core-shell structured microbubble comprising a fluorescent label; injecting a predetermined amount of the emulsion of the plurality of core-shell structured microbubbles intravenously to a body of a patient; tracking the fluorescent label of each respective core-shell structured microbubble utilizing a fluorescent detector for a time period of maximum 72 hours; and applying an ultrasonic energy to chest of the patient with an acoustic intensity in a range of 1 mWcm”2 to 600 mWcm”2.
11. The method of claim 10, wherein injecting the predetermined amount of the emulsion of the plurality of core-shell structured microbubbles intravenously to the body of the patient comprises injecting an amount of the emulsion of the plurality of core-shell structured microbubbles in a range of 10 pg/mL to 500 pg/mL intravenously to the body of the patient.
12. The method of claim 10, wherein applying the ultrasonic energy to the chest of the patient comprises applying the ultrasonic energy with an acoustic frequency in a range of 1
MHz to 2 MHz to the chest of the patient.
13. The method of claim 10, wherein applying the ultrasonic energy to the chest of the patient comprises applying the ultrasonic energy to the chest of the patient after reaching the drug delivery composition to the target place, wherein the target place comprises platelet-clogged arteries that supply blood to heart.
14. The method of claim 10, wherein applying the ultrasonic energy comprises applying the ultrasonic energy to the chest of the patient for a time period in a range of 1 minutes to 30 minutes.
15. The method of claim 10, wherein forming the emulsion of the plurality of core-shell structured microbubbles comprises: forming a shell portion, comprising: forming poly(D,L-lactide-co-glycolide)-NHS (PLGA-NHS) by dissolving PLGA-COOH, l-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and NHS in an anhydrous dichloromethane solution; forming PLGA-NH2 by dissolving the formed PLGA-NHS and ethylenediamine (EDA) in a dichloromethane solution; forming PLGA-HP by mixing dissolved heparin salt, NHS, and EDC with the formed PLGA-NH2 in N,N-dimethylformamide/formamide (DMF/FA); forming HOOC-PEG-COOH by dissolving HO-PEG-OH, succinic anhydride, and 4-(dimethylamino) pyridine (DMAP) in a dichloromethane solution; forming HOOC-PEG-NHS by dissolving the formed
HOOC-PEG-COOH, EDC, and NHS in an anhydrous dichloromethane solution; forming PLGA-HP-PEG-NHS by mixing the formed
HOOC-PEG-NHS, the formed PLGA-HP, DMAP, and dicyclohexylcarbodiimide (DCC) together; forming PLGA-HP-PEG-cRGD by mixing the formed
PLGA-HP-PEG-NHS with a cyclic arginine-glycine-aspartate (cRGD) solution; forming PLGA-HP-PEG-cRGD-platelet by mixing a platelet solution with the formed PLGA-HP-PEG-cRGD; forming a basic fibroblast growth factor (bFGF) loaded PLGA-HP- PEG-cRGD-platelet solution by dissolving the formed PLGA-HP-PEG-cRGD- platelet with bFGF in a phosphate -buffered saline (PBS) solution; forming a stabilized bFGF loaded PLGA-HP-PEG-cRGD-platelet solution by adding at least one stabilizing agent to the bFGF loaded PLGA-HP- PEG-cRGD-platelet solution; fluorescent labeling the bFGF loaded PLGA-HP-PEG-cRGD-platelet by mixing at least one fluorescent label with the bFGF loaded PLGA-HP-PEG- cRGD-platelet solution; and injecting the fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD- platelet solution into a polyvinyl alcohol (PVA) solution using a first syringe pump; and forming a core portion, comprising: injecting liquid perfluorohexane (PFH) into the PVA solution using a second syringe pump.
16. The method of claim 15, wherein forming PLGA-HP comprises mixing dissolved heparin salt, NHS, and EDC with the formed PLGA-NH2 in DMF/FA at a temperature in a range of 4°C to 25°C for a time period in a range of 100 minutes to 2000 minutes.
17. The method of claim 15, wherein injecting the fluorescent labeled bFGF loaded PEGA- HP-PEG-cRGD-platelet solution into the PVA solution comprises injecting the fluorescent labeled bFGF loaded PLGA-HP-PEG-cRGD-platelet solution with a flow rate in a range of 100 pl/min to 500 pl/min into the PVA solution using the first syringe pump.
18. The method of claim 15, wherein injecting the liquid PFH into the PVA solution comprises injecting the liquid PFH into the PVA solution with a flow rate in a range of 1 pl/min to 500 pl/min using the second syringe pump.
19. The method of claim 15, wherein forming the PLGA-HP-PEG-cRGD-platelet comprises mixing the platelet solution with the PLGA-HP-PEG-cRGD with a weight ratio in a range of 100:1 to 10:1 (PLGA-HP-PEG-cRGD: platelet).
20. The method of claim 15, wherein forming the PLGA-HP-PEG-cRGD comprises mixing the formed PLGA-HP-PEG-NHS with the cRGD solution with a weight ratio in a range of 10000:1 to 100:1 (PLGA-HP-PEG-NHS: cRGD).
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002036161A2 (en) * 2000-10-30 2002-05-10 Imarx Therapeutics, Inc. Novel targeted compositions for diagnostic and therapeutic use
US20040213756A1 (en) * 2003-04-15 2004-10-28 Michal Eugene T. Methods and compositions to treat myocardial conditions

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002036161A2 (en) * 2000-10-30 2002-05-10 Imarx Therapeutics, Inc. Novel targeted compositions for diagnostic and therapeutic use
US20040213756A1 (en) * 2003-04-15 2004-10-28 Michal Eugene T. Methods and compositions to treat myocardial conditions

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
DURO-CASTANO ET AL.: "Modulating angiogenesis with integrin-targeted nanomedicines", ADVANCED DRUG DELIVERY REVIEWS, vol. 119, 15 September 2017 (2017-09-15), pages 101 - 119, XP085268736, DOI: 10.1016/j.addr.2017.05.008 *
FIGUEIREDO, MARXA; ESENALIEV, RINAT: "PLGA Nanoparticles for Ultrasound-Mediated GeneDelivery to Solid Tumors ", JOURNAL OF DRUG DELIVERY, vol. 2012, no. Special Issue: Nanoparticles for Targeted Delivery, 28 February 2012 (2012-02-28), pages 767839, 1 - 767839, 20, XP009546793, ISSN: 2090-3014, DOI: 10.1155/2012/767839 *
HUANG YU, GU BORAM, SALLES-CRAWLEY ISABELLE I., TAYLOR KIRK A., YU LI, REN JIE, LIU XUHAN, EMERSON MICHAEL, LONGSTAFF COLIN, HUGHE: "Fibrinogen-mimicking, multiarm nanovesicles for human thrombus-specific delivery of tissue plasminogen activator and targeted thrombolytic therapy", SCIENCE ADVANCES, vol. 7, no. 23, 4 June 2021 (2021-06-04), XP093070422, DOI: 10.1126/sciadv.abf9033 *
HUANG YU, YU LI, REN JIE, GU BORAM, LONGSTAFF COLIN, HUGHES ALUN D., THOM SIMON A., XU XIAO YUN, CHEN RONGJUN: "An activated-platelet-sensitive nanocarrier enables targeted delivery of tissue plasminogen activator for effective thrombolytic therapy", JOURNAL OF CONTROLLED RELEASE, ELSEVIER, AMSTERDAM, NL, vol. 300, 1 April 2019 (2019-04-01), AMSTERDAM, NL , pages 1 - 12, XP093070420, ISSN: 0168-3659, DOI: 10.1016/j.jconrel.2019.02.033 *
LIANG Y ET AL.: "Heparin-functionalized polymeric biomaterials in tissue engineering and drug delivery applications", ACTA BIOMATERIALIA, vol. 10, no. 4, 2 August 2013 (2013-08-02), pages 1588 - 600, XP055736568, DOI: 10.1016/j.actbio.2013.07.031 *
MODERY PAWLOWSKI CHRISTA, KUO HSIAO-HSUAN, BALDWIN WILLIAM, GUPTA ANIRBAN: "A platelet-inspired paradigm for nanomedicine targeted to multiple diseases", NANOMEDICINE, FUTURE MEDICINE LTD., LONDON, GB, vol. 8, no. 10, 30 September 2013 (2013-09-30), GB , pages 1709 - 1727, XP009546030, ISSN: 1743-5889, DOI: 10.2217/nnm.13.113 *
SHEN ET AL.: "Cell affinity for bFGF immobilized heparin-containing poly(lactide-co-glycolide) scaffolds", BIOMATERIALS, vol. 32, no. 13, May 2011 (2011-05-01), pages 3404 - 12, XP028366484, DOI: 10.1016/j.biomaterials.2011.01.037 *
XUAN JIQING, CHEN YULI, ZHU LEILEI, GUO YUAN, DENG LIMING, ZHENG YUANYI, WANG ZHAOXIA, WANG ZHIGANG, AO MENG: "Ultrasound molecular imaging with cRGD-PLGA-PFOB nanoparticles for liver fibrosis staging in a rat model", ONCOTARGET, vol. 8, no. 65, 12 December 2017 (2017-12-12), pages 108676 - 108691, XP093070423, DOI: 10.18632/oncotarget.21358 *

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