Paper page

Original: publisher page

Ming Lu, Lynne S. Taylor* · Crystal Growth & Design (Just Accepted Manuscript, published on the web 29 Aug 2016) (2016)

Vemurafenib: A Tetramorphic System Displaying Concomitant Crystallization from the Supercooled Liquid

Glassy vemurafenib, heated above its Tg of 108 °C, crystallizes into two or three of four polymorphs at once (the known α and the new β, γ and δ), and which form wins at each temperature follows how fast each form nucleates and grows, not which form is most stable.

Abbreviations

ASD amorphous solid dispersion; DSC differential scanning calorimetry; FTIR Fourier transform infrared spectroscopy (ATR mode); HSM hot-stage microscopy; POM polarized light (optical) microscopy; PXRD (XRPD) powder X-ray diffraction; Tg glass transition temperature; Tm melting temperature; Tcc cold-crystallization (isothermal crystallization) temperature; ΔHm enthalpy of fusion; ΔHc crystallization enthalpy; VMN vemurafenib (Fig. 6 label); p. = printed manuscript page; SI p. = page of the supporting information.

Graphical abstract: Six crystal types from one supercooled liquid
Graphical abstract · p.27 · Six crystal types from one supercooled liquidShows. the vemurafenib structure with arrows to α-needle (200–270 °C), α-diamond spherulite (170–200 °C), α-brown feather (150–240 °C), β (110–240 °C), γ (170–190 °C) and δ (110–165 °C), and the conversions δ to β (170–225 °C), γ to β (200–220 °C) and β to α (150–240 °C).

Story. Why they did it → how → what they found → how it fits together → what is left. Each part opens with the short version; "More detail" under it gives the full account. Grey ids open the evidence behind a sentence.

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Question

Which crystal forms grow when amorphous vemurafenib crystallizes from its supercooled liquid between Tg and Tm, how fast does each grow, how do they convert into one another, and how are they related thermodynamically?

Short answer

Four polymorphs in six morphologies: α (needle, diamond spherulite, brown feather), β, γ and δ. Isothermal crystallization at 120–240 °C typically gave two or three forms together, mostly β or α; above about 250 °C only α grew. β grew fastest up to 230 °C (≈110 µm/s) and dominated at 150–190 °C, although α is the stable form and is monotropic to β. ····

Conclusion

For melt-quenched vemurafenib crystallized between its Tg and Tm, the authors conclude that three new metastable polymorphs (β, γ, δ) form alongside the known α, and that "it thus appears that vemurafenib is at least a tetramorphic system" with a high tendency for concomitant polymorphism. · They rank stability as α > β > γ or δ: γ and δ convert to β, and β converts to α, slowly in the solid state where it touches α and, near 250 °C, through the melt. ·· α and β are monotropic, "inferred" from the heat-of-fusion rule and supported by a Hoffman estimate with a virtual transition at 319 °C. · Because β dominates from 120 to 190 °C despite the larger driving force for α, "these observations indicate" that cold crystallization is controlled by factors other than the thermodynamic driving force, which the authors explain by different nucleation and growth zones for each form. · The nucleation side of that explanation was judged from microscope impressions rather than measured, γ and δ were not obtained pure, and the γ–δ stability relationship "could not be established". ··

01Why this study

Known 1Amorphous drugs tend to crystallize

They dissolve faster than crystals but revert to crystals in storage or dissolution and lose that advantage.

Loftsson 2010, ref 1

Known 2Cold crystallization is a distinct route

Crystallization on heating a glass into its supercooled liquid is well studied in polymers, much less in drugs.

Nagarajan 2015, ref 2; Wellen 2005, ref 4

Known 3Few studies follow which crystal forms an amorphous drug produces

Work has focused on mobility, polymers and water, not on phase evolution.

Purohit 2015, ref 9; refs 5–8 · refs 10–16

Known 4Vemurafenib grows crystals very fast

Its maximum crystal velocity is 106 ± 4 µm/s at 250 °C, yet the marketed product is an ASD.

Trasi 2014, ref 18 ··

Gap → this paperHow does amorphous vemurafenib crystallize across the temperature range from Tg to Tm, and which polymorphs does it produce?
More detail

What was already known. The introduction builds on these points:

  • Poor solubility and the amorphous route. By the biopharmaceutics classification system, 90% of development compounds and 40% of marketed drugs are poorly water-soluble (Loftsson 2010, ref 1). Amorphous forms lack a crystal lattice, which lowers the energy barrier to dissolution, but they are thermodynamically un/metastable and recrystallize during storage or dissolution, losing the amorphous solubility advantage. Inhibiting recrystallization, and so understanding crystallization kinetics, is therefore important.
  • Melt versus cold crystallization. Melt crystallization happens on cooling the melt; cold crystallization on heating the glass or supercooled liquid. Cold crystallization is well studied in polymers, where amorphous chain segments rearrange into a crystal (Nagarajan 2015, ref 2; Zhou 2016, ref 3; Wellen 2005, ref 4); in small molecules the molecules rearrange into the lattice. Below Tg the material is a glass; between Tg and Tm it is a supercooled liquid.
  • What earlier work covered. Research on amorphous pharmaceuticals has examined molecular mobility (Sibik 2016, ref 5; Mehta 2016, ref 6; Yu 2016, ref 7), polymeric additives (Tian 2015, ref 8) and water (Purohit 2015, ref 9). How different crystal phases evolve from an amorphous drug has been studied rarely (refs 10–16), including Yu's work on concomitant polymorphs and cross-nucleation (refs 13–16).
  • Vemurafenib. It is practically insoluble in water (below 0.1 µg/ml; Shah 2013, ref 17). Preliminary studies found a strong crystallization tendency and a maximum crystal velocity of 106 ± 4 µm/s at 250 °C, fast compared with other compounds (Trasi 2014, ref 18), so the amorphous form "might be anticipated" to be physically unstable; the commercial formulation nevertheless contains the drug as an amorphous solid dispersion (no reference given). ···

What the paper set out to do. Measure the crystallization kinetics of amorphous vemurafenib as a function of temperature and map its polymorphic landscape.

02How they approached it

One drug and no excipient: make a glass by quenching the melt, then crystallize the supercooled liquid at fixed temperatures from just above Tg (120 °C) to just below Tm (270 °C), watching each crystal grow on a hot stage and identifying it on the spot by Raman microscopy. ···

Amorphous vemurafenibmelt-quenched glassStarting material for all cold crystallization; Tg 108 °C; measured. ·
α-formas-received form; needle, diamond spherulite and brown feather morphologiesHighest melting point, 271 °C, and 114 J/g; most stable; measured. ·
β-formnew; spherulitesMelts at 248 °C with 68 J/g; fastest-growing form below 230 °C; measured. ·
γ-formnew; small spherulitesNo melting point, converts to β first; identified by Raman; measured. ·
δ-formnew; brown clustersNo melting point, converts to β first; Raman and a partial PXRD pattern; measured. ·
Patent Forms A and Bliterature solvate and desolvate, ref 26PXRD peaks and melting points compared only; cited.

What was done

Experiment

As-received α melted at 285 °C for 1 min→quenched on a metal block→glass, no degradation by HPLC.·

↓
Experiment

DSC of crystalline α (heat, cool, reheat at 10 °C/min) and of the glass heated at 10 °C/min→hot-stage view and PXRD of samples quenched at 170 and 236 °C.····

↓
Experiment

Isothermal crystallization of the supercooled liquid at 120–270 °C on a hot stage→morphology and growth rate of each crystal type.··

Experiment

Raman microscopy of each morphology, PXRD of isothermal samples, ATR-FTIR of amorphous, α and β→form identity.·····

↓
Experiment

Heating partly crystallized samples at 10 °C/min (γ from 190 °C, δ from 150 °C), holds at 170, 190 and 230 °C, DSC at 100 °C/min→transformation paths.·····

↓
Calculation

Tm and ΔHm of α and β→heat-of-fusion rule and Hoffman equation→free energy against temperature, virtual transition at 319 °C.···

More detail

What was done

  1. Glass. The as-received α powder (>99%, Attix Pharmaceuticals), placed between two coverslips, was held at 285 °C for 1 min and quenched on a metal block. The glass was clear and amorphous under the polarizing microscope, did not change colour and showed no degradation by HPLC. Slow cooling (10 °C/min) did not give a glass. ··
  2. DSC. TA Q2000 at 10 °C/min: the as-received crystal was heated, cooled and reheated; the glass, quenched in the DSC pan, was heated from −40 to 285 °C. A second glass was annealed at 150 °C for 20 min to grow β and then heated at 100 °C/min to see β melting. The α and β samples of Fig. 12 were made by cooling the melt at 10 °C/min (α) and by holding at 190 °C for 20 min after melting at 280 °C (β). The methods and the Fig. 2 caption give different end temperatures and holds. ··
  3. Isothermal cold crystallization. Glass films on a Linkam hot stage were held at a fixed Tcc between 120 °C (just above Tg) and 270 °C (just below Tm) under a polarizing microscope with time-lapse photography. Crystal size was measured on the images and the growth rate taken as the slope of size against time. The number of crystals per temperature and the route to Tcc are not stated. ··
  4. Identification. Raman microscopy (532 nm, 2.1 µm spot, 10× objective) gave a spectrum from each spherulite or crystal type; PXRD (Cu Kα, 4.5–30° 2θ) was run on isothermal samples, on samples quenched during heating and on α and β references; ATR-FTIR could be run only on amorphous, α and β, the forms available in enough quantity. ···
  5. Transformations. A sample crystallized at 190 °C (α, β, γ) and a δ-only area grown at 150 °C were heated at 10 °C/min under the microscope, with Raman of each region; the β-to-α front was followed at 170 and 230 °C and the γ-to-β change during a hold at 190 °C; a solvent-mediated conversion at room temperature was run but not shown. ······
  6. Thermodynamics. The heat-of-fusion rule (Burger–Ramberger, ref 28) was applied to α (271 °C, 53.8 kJ/mol) and β (248 °C, 33.3 kJ/mol), and the Hoffman equation (ref 29) gave free-energy curves of both forms against temperature. ··

Key equations

Hoffman equation (Eq. 1)
ΔGv=ΔHm(Tm−T)TTm2

An estimate of the free-energy gain when the liquid crystallizes at temperature T, from the melting point and heat of fusion alone. With α (271 °C, 53.8 kJ/mol) and β (248 °C, 33.3 kJ/mol) it gives α the larger driving force at every temperature up to the virtual transition at 319 °C, above both melting points.·

Naito relation (in words, p.13)
Tmax≈0.94Tm

The fastest crystal growth is expected near 0.94 of the melting temperature in kelvin; for α this gives 240 °C, against the 250 °C observed.·

Terms used on this page
  • Cold crystallization: crystallization on heating a glass above its Tg, from the supercooled liquid; melt crystallization happens on cooling the melt.
  • Supercooled liquid: the amorphous state between Tg and Tm; mobile, but below the melting point.
  • Concomitant polymorphs: two or more crystal forms that crystallize at the same time from the same liquid.
  • Spherulite: a round aggregate of crystal fibres radiating from a centre; needle: an elongated crystal.
  • Monotropic: one form is the more stable one at all temperatures below melting; enantiotropic: the order switches at a transition temperature below the melting points.
  • Heat-of-fusion rule: if the higher-melting form also has the larger heat of fusion, the two forms are monotropic.
  • Virtual transition temperature: where the free-energy curves of two monotropic forms would cross; it lies above both melting points and is never reached.
  • Solid–solid versus melt-mediated transformation: conversion inside the solid versus melting of one form followed by crystallization of the other.

03Main findings

1

The melt crystallizes readily on cooling; the glass crystallizes on heating in several steps

Cooled at 10 °C/min, the melt crystallized to α from 248.9 °C, only about 25 °C below its melting point (271.3 °C). The quenched glass (Tg 108 °C) showed a sharp exotherm at 157 °C, a broad one at 200–237 °C, a small endotherm–exotherm pair at 250 °C and α melting at 271.5 °C. ··

Fig. 3: Heating the glass: DSC with hot-stage images, and PXRD
Fig. 3 · p.7 · Heating the glass: DSC with hot-stage images, and PXRDShows. exotherm at 157 °C (β, with δ in yellow rings), broad exotherm at 217 °C (β to α, δ to β), small pair at 250 °C (inset), α melting at 271.5 °C; PXRD at 170 °C (β) and 236 °C (β plus α).
More detail

The as-received powder melted sharply at 271.3 °C (ΔHm 55.9 kJ/mol). On cooling at 10 °C/min it crystallized from 248.9 °C (exotherm peak 246.7 °C, ΔHc 48.7 kJ/mol) and on reheating melted again at 271.5 °C as α, confirmed by XRPD. With only about 25 °C of supercooling before crystallization, the authors call vemurafenib a "relatively poor glass former" ("suggesting"); a glass forms only on faster cooling. ··

The glass heated at 10 °C/min (Fig. 3A) passed its Tg at 108 °C, then gave a sharp exotherm from about 152 °C (peak 157.2 °C, ΔHc 23.6 kJ/mol), a broad exotherm from about 200 °C (peak 217.3 °C, ending at 237 °C, ΔH 19.6 kJ/mol), a very small endotherm at 249.6 °C immediately followed by an exotherm at 250.5 °C, and a final melting peak at 271.5 °C whose ΔHm is given as 51.0 kJ/mol on p.6 and 53.8 kJ/mol on p.18. Hot-stage images of the same heating show mainly β spherulites with a few δ after the first exotherm, α-brown feathers spreading over β from 200 °C, residual β melting at 250 °C and all α crystals melted by 277 °C. PXRD of samples quenched at 170 °C showed β; at 236 °C α peaks had appeared while several β peaks remained. ····

The authors read the first exotherm as crystallization to "predominantly" β with a few δ; the broad exotherm as the slow, exothermic solid–solid β-to-α change, incomplete at 236 °C and occurring at the surface of β crystals; the 250 °C pair as melting of residual β offset by crystallization of α-needles ("it is likely"); and the last peak as α melting. ···

Fig. 2: The melt crystallizes to α on cooling
Fig. 2 · p.6 · The melt crystallizes to α on coolingShows. α melts at 271.3 °C, the melt crystallizes on cooling with a peak at 246.7 °C, and the product melts again as α at 271.5 °C.
2

Four polymorphs in six morphologies

Hot-stage microscopy and Raman spectra separated α (as needles, diamond spherulites and brown feathers, all with one Raman spectrum and Tm 271 °C), β (spherulites, Tm 248 °C), γ (small spherulites) and δ (brown clusters); γ and δ convert to β before they melt. ···

Fig. 4: The six morphologies and their Raman spectra
Fig. 4 · p.8 · The six morphologies and their Raman spectraShows. A–C the three α morphologies with identical spectra, D β, E γ, F δ, each with its growth region and melting or transformation temperature.Read it as. the letters printed in the figure (A needle, B diamond spherulite, C brown feather), not those in the caption.Also supports K1
More detail

The Fig. 4 catalogue, by the panel letters in the figure: A α-needles (growth region 200–270 °C), B α-diamond spherulites (170–200 °C), C α-brown feathers (150–240 °C), all melting at 271 °C; D β spherulites (110–210 °C, melting at 248 °C); E γ spherulites (170–200 °C, turning into β at 200–225 °C); F δ dense brown clusters (110–165 °C, turning into β at 170–225 °C). Fig. 5 and the text give somewhat different ranges, and the caption and text letter the panels differently. ···

The three α morphologies gave identical Raman spectra, also side by side at 200 °C, and the same melting point, which the authors read as one crystal structure ("suggesting", later "confirming"). The other morphologies differed chiefly between 1550 and 1700 cm−1: ring C=C bands at 1597–1613 cm−1, and C=O bands at 1640 cm−1 (α), 1640 and 1655 cm−1 (β) and 1624 cm−1 (δ); γ showed bands at 1600 and 1610 cm−1. The authors take the C=O differences as "suggesting" different C=O···H–N interactions in the crystal structures, and the Raman spectra as "strong evidence" for a tetramorphic system. ····

PXRD separates α (peaks from 6.8 to 24.4° 2θ; 114 J/g) from β (5.0, 8.4, 9.9, 12.0, 14.9, 16.7, 20.8 and 27.2°; 68 J/g). δ-specific peaks were reported at 8.8, 11.4, 11.7 and 16.3° from a β + δ sample made at 120 °C; γ was too minor to see by PXRD; no PXRD pattern, Tm or ΔHm is given for γ or δ. The as-received α matches the crystalline form reported by Shah et al.; two patent forms (A, a probable solvate, and B, its desolvate) were judged not to match any form here. ·······

3

Which forms appear depends on the crystallization temperature

At 120 °C β and δ nucleate; at 170–190 °C β, γ and α-diamond spherulites crystallize together; above 200 °C α (as needles above 210 °C) takes over from β; above 250 °C only α grows. β dominates from 150 to 190 °C. ··

Fig. 5: Where each form grows and where it converts
Fig. 5 · p.9 · Where each form grows and where it convertsShows. growth ranges of each form on a Tg–Tm axis, with arrows δ to β, γ to β and β to α.Also supports K4
More detail
  • 120–165 °C. At 120 °C only β and δ nucleate, with similar growth rates, and the PXRD pattern carries β and δ peaks. δ nucleates densely at low temperature but is engulfed by β; no δ was seen at 165 °C and no α at 165 °C or below. β dominates from 150 °C. ·
  • 170–190 °C. β still dominates; α-diamond spherulites (170–200 °C) are always embedded in β; γ nucleates (170–190 °C) and stays a minor fraction surrounded by β. PXRD at 190 °C shows β. ·
  • 200–240 °C. α-needles appear at 200 °C and above 210 °C α grows only as needles; β nucleates less readily beyond 200 °C and erratically, with low frequency, at 220–240 °C. PXRD at 230 °C shows α and β together. ·
  • Above 250 °C. β no longer nucleates and no brown feather forms; PXRD at 260 °C matches the as-received α. ·

The abstract summarizes this as two or three of the four polymorphs in each sample at 120–240 °C, α or β typically dominating, and only α at higher temperature; the text implies a single form at 165 °C. ·

Fig. 6: PXRD after isothermal crystallization
Fig. 6 · p.10 · PXRD after isothermal crystallizationShows. (A) α at 260 °C, α plus β at 230 °C, β at 130–190 °C, β plus δ at 120 °C; (B) α and β reference patterns.Also supports K2
4

Growth rates differ by up to two orders of magnitude

β grew fastest up to 230 °C (≈12 µm/s at 170 °C, ≈69 at 190 °C, ≈110 at 230 °C); α-needles rose from ≈41 µm/s at 210 °C to a maximum of ≈106 µm/s at 250 °C; α-diamond spherulites reached ≈21 µm/s at 200 °C; γ, δ and brown feathers grew at ≈4 µm/s or less.

Fig. 7: Growth rate against temperature for every form
Fig. 7 · p.11 · Growth rate against temperature for every formShows. β (blue) fastest up to 230 °C; α-needles (purple) bell-shaped with the maximum at 250 °C; α-diamond spherulites (red) slower; γ, δ and brown feathers near zero.Also supports K5, K6
More detail

Read from Fig. 7: β was ≈0 at 120–150 °C, ≈5–6 µm/s at 155–160 °C and ≈12–14 µm/s at 170–180 °C, then jumped to ≈69 µm/s at 190 °C (error bar ≈45–94) and reached ≈94, ≈99 and ≈110 µm/s at 210, 220 and 230 °C. α-needles grew at ≈41, 60, 85, 98, 106 and 92 µm/s from 210 to 260 °C and ≈14 µm/s near 270 °C: a bell-shaped curve. α-diamond spherulites grew at ≈1, 3, 11 and 21 µm/s at 170, 180, 190 and 200 °C, on the same temperature trend as the needles. γ reached ≈4 µm/s at 190 °C, δ stayed near zero at 120–165 °C, and α-brown feathers stayed at or below ≈1 µm/s, with 0.3 ± 0.1 µm/s measured on β at 220 °C. The error-bar type and number of crystals are not stated. ··

The authors explain the bell shape by a driving force that rises and a molecular mobility that falls as temperature drops ("can be readily explained"), note that Naito's 0.94 Tm rule puts the maximum at 240 °C against the observed 250 °C, and call the ≈106 µm/s maximum very fast compared with other drug-like molecules ("it is apparent"). ····

5

The metastable forms convert step by step: γ and δ to β, β to α

γ converted to β above 200 °C on heating (complete near 220 °C) and within 60 min at 190 °C; δ converted to β at about 220 °C; β converted to α slowly in the solid state, only where it touched α, and quickly after β melted at about 250 °C. ·····

Fig. 8: β turning into α at 170 °C
Fig. 8 · p.11 · β turning into α at 170 °CShows. a brown α front spreading from a diamond spherulite into β at 5, 10 and 25 min; Raman of I and II matches α, III is β; α peaks appear in PXRD by 60 min.Also supports K7
More detail
  • γ to β. In a sample crystallized at 190 °C (α, β, γ) and heated at 10 °C/min, γ turned into a new phase above 200 °C, complete around 220 °C; that phase melted together with the original β at 254–255 °C and gave a β Raman spectrum. Held at 190 °C, the same conversion needed 60 min. ···
  • δ to β. A small δ-only area grown at 150 °C and heated at 10 °C/min was swept by a β front between 219 and 225 °C. No DSC peak was seen for this step, "probably" because little δ was present. ··
  • β to α in the solid state. At 170 °C a dark brown α front ("brown feather") grew from a diamond spherulite into the surrounding β within 5–25 min; Raman of the front matched α, and PXRD after 60 min showed α peaks added to β. The same front appeared at 230 °C. It always started at an α:β interface, and β without α contact did not transform. Brown feathers grew at 0.3 ± 0.1 µm/s at 220 °C, "presumably" because of a high activation barrier. ······
  • β to α through the melt. On heating, β melted at about 250 °C and α-needles grew from the edges of α-spherulites (Fig. 9D, E); at 100 °C/min after a 150 °C anneal, the β melting peak (≈252 °C) was followed at once by a recrystallization exotherm and α melting. The separate Fig. 12 β sample showed no recrystallization after melting. ····
  • At room temperature. β converted to α in a solvent; the data are not shown.
Fig. 9: γ turning into β on heating, then α-needles growing as β melts
Fig. 9 · p.14 · γ turning into β on heating, then α-needles growing as β meltsShows. γ (III) becomes β (IV) between 190 and 222 °C; α-needles (V) grow at 253–254 °C; Raman of I–IV.Also supports K4
Fig. 10: δ turning into β on heating
Fig. 10 · p.15 · δ turning into β on heatingShows. a β front sweeping through δ between 219 and 225 °C; Raman of δ (I) and β (II).Also supports K4
6

α is the stable form and is monotropically related to β

α melts higher (271 °C) and with a larger heat of fusion (53.8 kJ/mol) than β (248 °C, 33.3 kJ/mol), so by the heat-of-fusion rule the pair is monotropic; the Hoffman equation gives α the lower free energy at all temperatures up to a virtual transition at 319 °C. ···

Fig. 13: Hoffman free energy of α and β
Fig. 13 · p.23 · Hoffman free energy of α and βShows. α lies below β at every temperature below its melting point; the curves would cross near 319 °C.Read it as. crystal minus liquid, so values are negative below Tm.Also supports K3
More detail

DSC (Fig. 12) shows α melting at 271.5 °C and β at 247.9 °C; the text gives Tm 271 and 248 °C with ΔHm 53.8 and 33.3 kJ/mol (Table 2: 114 and 68 J/g). The heat-of-fusion rule (ref 28) then makes the pair monotropic, which the authors find "consistent with" the exothermic β-to-α change and with β always converting to α between 25 and 250 °C. The Hoffman curves (Eq. 1) put α below β at every temperature studied, ≈−11.0 against ≈−6.3 kJ/mol relative to the liquid near 115 °C (Fig. 13 plots the negative of ΔGv), and cross only at a virtual transition of 319 °C (recomputed 318.7 °C), above both melting points. The abstract gives the overall stability order as α > β > γ or δ. ··········

Fig. 12: Melting of α and β
Fig. 12 · p.23 · Melting of α and βShows. α melts at 271.5 °C, β at 247.9 °C.Also supports K3

04Further observations

  • The as-received α matches the crystalline vemurafenib described by Shah et al.
  • Two patent forms (A and B) were judged different from all four forms. ·
  • α-needles have a smoother surface and a sharper interface with the melt than α-spherulites.
  • Diamond spherulites look shiny only when formed at 170–180 °C. ·
  • FTIR was possible only for amorphous, α and β; C=O bands are stronger in IR, ring bands in Raman. ·
More detail
  • Known form. The diffractogram of the α-form is the same as that of the crystalline vemurafenib reported by Shah et al. (ref 17); the comparison pattern is not shown.
  • Patent forms. A patent describes Form A, made from solvent and judged a solvate, and Form B, made by drying Form A. Their PXRD peaks (A: 6.5, 9.7, 18.0, 19.5, 23.7°; B: 5.9, 9.1, 14.2, 18.3, 19.7, 23.7°) and melting points (268.5 and 270.9 °C) were judged not to match any form here, although Form B melts within 0.1 °C of α and shares four peaks with it within 0.2°. ··
  • Morphology and undercooling. α grew as spherulites at 170–200 °C and as smooth, sharply bounded needles from 200 °C; the authors cite Jackson (ref 19), for whom faceted or spiky crystals form at small undercooling and spherulites at large undercooling through kinetic roughening. ··
  • Diamond spherulites. They were shiny when formed at 170–180 °C, were always embedded in β, and grew larger at higher temperature before the liquid was used up. ··
  • All α morphologies stay α. None of them changed form before melting.
  • Spectra. ATR-FTIR could be measured only for the amorphous form, α and β, the forms available in enough quantity. Ring C=C stretching appears at 1589–1613 cm−1 and C=O stretching at 1640 cm−1 in both spectra; C=O bands are stronger in the IR and ring bands in the Raman spectrum. ·

05How the authors connect the findings

1Each polymorph has its own temperature zone for nucleation and its own zone for growth.
↓
2At a given temperature the overlap of those zones and the size of the two rates decide which forms appear and how much of each, whatever the stability order. ·
↓
3The metastable products then move toward α: γ and δ turn into β, and β into α at α contacts or after it melts. ··
↓
Low temperature (120–165 °C)β and δ nucleate↓δ nucleates densely but grows extremely slowly↓β overgrows δ and dominates from 150 °C.··
Middle (170–190 °C)β nucleates sparsely but grows fastest↓γ and α-spherulites nucleate but grow slowly↓β dominates, γ stays minor.··
High (above 200 °C)β nucleation becomes unfavourable↓α nucleates readily and grows fast↓α predominates; above 250 °C only α.··
What sets the switch point. The temperature at which β stops nucleating readily (above about 200 °C) while α nucleates from about 170 °C and grows fast; nucleation was judged by eye, not measured. ··
Conclusion drawn. Cold crystallization of amorphous vemurafenib is "controlled by factors other than the thermodynamic driving force" ("These observations indicate"), in line with metastable forms being more probable at large undercooling. ·
More detail

The authors start from three ideas in the literature: Ostwald's law of stages, in which the least stable form crystallizes first and converts stepwise (ref 22); the view that concomitant polymorphs have similar nucleation rates (refs 23–25); and Yu's point that a fast-nucleating but slow-growing form can be overtaken by a slow-nucleating, fast-growing one, together with cross-nucleation of one form on another (refs 13, 15, 16).

  1. Different polymorphs have different nucleation and growth zones (Woo, ref 27); their overlap and magnitudes at the crystallization temperature set which forms appear and in what amount ("This can be explained").
  2. α nucleates fast enough near its growth maximum (250 °C) to crystallize on cooling the melt; since the nucleation maximum lies below the growth maximum (Okui, ref 20) and α never appears at 165 °C or below, its favourable nucleation zone is "approximately" 170–270 °C ("it appears").

Low temperature (120–165 °C). δ "exhibits a fast nucleation rate" and high nucleation density, but grows extremely slowly and is engulfed by β; at 120 °C only β and δ nucleate, with similar growth rates. ·

Middle (170–190 °C). Nucleation "appears to be the limiting step" for β: few β nuclei form above 150 °C, but they grow fast into large regions below 200 °C. γ is "an example of a polymorph that nucleates rapidly but grows slowly", so it stays a low mass fraction at 170–190 °C. On heating the glass at 10 °C/min the crystallization peak near 160 °C falls where β grows fastest, so β "would be expected" to dominate the DSC run. ···

High (above 200 °C). β nucleation becomes less favourable; although β still grows faster than α, α "starts to be the predominant form due to more favorable nucleation".

  1. Only the α–β pair could be analysed thermodynamically, because γ and δ could not be obtained pure. The heat-of-fusion rule and the Hoffman equation give α the larger driving force at every temperature, yet β dominates between 120 and 190 °C, so the outcome is set by "factors other than the thermodynamic driving force"; this fits the literature observation that metastable forms are more probable at larger undercooling (Ishihara, ref 30). ····

06What it means for formulation

Stated by the authors
  • Amorphous drugs need crystallization control, which requires understanding their crystallization kinetics.
  • The rich polymorphic landscape of the supercooled liquid "underscores the complexity" of evaluating crystallization of amorphous drug forms.
AI reading, not stated in the paper
  • When amorphous vemurafenib crystallizes above Tg, the first crystals are likely to be the metastable β (or δ close to Tg), not α; crystallinity checks should look for β, γ and δ signatures.
  • Heat exposure between about 120 and 240 °C, as in melt processing, can produce forms other than α in the pure drug.
More detail
Stated by the authors
  • Amorphous drugs raise solubility but recrystallize in storage or dissolution, so recrystallization must be inhibited and the crystallization kinetics understood.
  • The discovery of three new polymorphs from the supercooled liquid "highlights the rich polymorphic landscape" accessible this way and "underscores the complexity" of evaluating the crystallization of amorphous drug forms.
  • Given its fast crystal growth, amorphous vemurafenib "might be anticipated" to be physically unstable, yet the commercial product is an amorphous solid dispersion. ·
AI reading, not stated in the paper
  • The form that appears depends on temperature: in the pure drug, β (with δ near Tg) forms at 120–190 °C and α above 200 °C. A dispersion that crystallizes during hot processing or high-temperature stress could therefore contain metastable forms; this paper tests no polymer, so the temperature windows apply to the pure drug only.
  • γ was hard to see by PXRD and was found by Raman microscopy; a routine PXRD check for α peaks alone could miss metastable forms present in small amounts.
  • β, γ and δ were not characterized for solubility, so the loss of supersaturation potential when an amorphous vemurafenib product crystallizes may depend on which form appears.
  • Growth near 100 µm/s means a single nucleus can cross a 100 µm region of pure drug in about a second at 210–250 °C; this is why crystal growth, not only nucleation, matters for keeping the drug amorphous.

07Open questions

Raised by the authors
  • What is the stability relationship between γ and δ?
  • What are the melting points and thermodynamic data of γ and δ, which could not be obtained pure? ·
Not covered by this paper (AI reading)
  • Nucleation rates of each form were not measured.
  • No crystal structures were determined; γ and δ rest on Raman and morphology.
  • Crystallization below Tg, with polymer or water, was not tested.
More detail
Raised by the authors
  • The stability order of γ and δ "could not be established".
  • γ and δ convert to β before melting, so their melting points could not be measured, and pure samples were difficult to obtain. ··
Not covered by this paper (AI reading)
  • Nucleation was judged from microscope impressions; nucleation rates or densities of each form against temperature would test the kinetic explanation directly.
  • γ and δ have no PXRD pattern, melting data or crystal structure; the stated δ PXRD peaks cannot be checked on the printed 120 °C pattern. ·
  • Growth rates come without the number of crystals or error type.
  • Only the pure drug was studied, between 120 and 270 °C and in a few solvent-free thermal paths; crystallization of the glass below Tg, in water or in the polymer of the commercial dispersion was not examined.
  • The free-energy ranking rests on one set of α and β enthalpies; the α value differs between parts of the paper.

08How solid

AI assessment, not a statement made in the paper
Moderate

The distinct α and β forms and their monotropic relationship rest on DSC, PXRD, Raman and direct observation of β turning into α. γ and δ rest on morphology and Raman alone, nucleation was never measured, and several temperature ranges and enthalpies are stated inconsistently. ······

More detail
What holds
  • α and β are distinct by Raman, PXRD and DSC melting, and β turning into α was watched directly and checked by Raman and PXRD. ···
  • The three α morphologies share one Raman spectrum and melting point. ·
  • Growth rates cover the whole range from just above Tg to just below Tm, and the α maximum (≈106 µm/s at 250 °C) matches the earlier preliminary value of 106 ± 4 µm/s. ·
  • The 319 °C virtual transition reproduces from the printed inputs (318.7 °C).
What weakens it
  • γ and δ were never isolated pure and have no PXRD pattern, melting point or heat of fusion; the stated δ PXRD peaks cannot be checked on the printed 120 °C pattern. ·
  • Nucleation was judged by eye; the kinetic explanation rests on growth rates plus impressions of nucleation density.
  • Growth rates come without a stated n or error type; the β error bar at 190 °C spans ≈45–94 µm/s.
  • The α heat of fusion appears as 51.0, 53.8 and 55.9 kJ/mol; the 319 °C result holds only for 53.8.
  • Temperature ranges for the same form differ between Fig. 4, Fig. 5 and the text, and the Fig. 4 panel letters do not match the caption and text. ·
  • The β sample for DSC was made at 190 °C, where α and γ also form, and its melting may overlap recrystallization; the claimed immediate melt-mediated β-to-α step is absent from the Fig. 12 trace. ··
  • The conclusion orders γ and δ although the Fig. 5 caption says their relationship could not be established.

09Figure guide

Every figure and table of the paper, what it is there for, and what to check on the original. Key figures are shown with the story.

FigureWhat it does in the paperSupportsCheck (AI note)
Fig. 1 (p.3)Defines the molecule, including the C=O and N–H groups discussed for the Raman bands.0.2A schematic, not data.
Fig. 2 (p.6)key figureShows the melt crystallizing to α on cooling with only about 25 °C of supercooling.F1End temperatures differ from the methods.
Fig. 3 (p.7)key figureDSC of the heated glass with hot-stage insets, and PXRD of quenched samples: β first, then β to α.··F1Two ΔHm values are given for the final peak.
Fig. 4 (p.8)key figureCatalogue of the six morphologies with Raman spectra and temperature ranges.·F2, K1Caption and text letter the panels differently.
Fig. 5 (p.9)key figureSummary of the growth and transformation ranges of each form.F3, K4Ranges differ from Fig. 4 and the text.
Fig. 6 (p.10)key figurePXRD of isothermal samples and of α and β references: the temperature map of forms.·F3, K2The 120 °C pattern has unassigned and α-position peaks.
Fig. 7 (p.11)key figureGrowth rates of all forms against temperature, the kinetic core of the paper.F4, K5, K6Number of crystals and error type not stated.
Fig. 8 (p.11)key figureβ turning into α at 170 °C, by microscopy, Raman and PXRD.F5, K7
Fig. 9 (p.14)key figureγ turning into β on heating, and α-needles growing as β melts.F5, K4Different sample history from the heated glass it is used to explain.
Fig. 10 (p.15)key figureδ turning into β on heating near 220 °C.F5, K4Panels labelled A, B, B.
Fig. 11 (p.16)FTIR and Raman of amorphous, α and β, used to assign bands.F2Axes labelled wavelength for wavenumber.
Table 1 (p.17)Raman and IR peaks of all forms; the C=O bands differ between forms.F2, K1
Table 2 (p.20)PXRD peaks, Tm and ΔHm of α, β and the patent forms; none for γ and δ.F2, F6, K3Form B melts within 0.1 °C of α.
Fig. 12 (p.23)key figureMelting endotherms of α and β used for the heat-of-fusion rule.F6, K3No recrystallization after β melting; β purity not shown.·
Fig. 13 (p.23)key figureHoffman free energy of α and β: monotropic, virtual transition at 319 °C.F6, K3Plots the negative of ΔGv; result tied to 53.8 kJ/mol.·
More detail
FigureWhat it does in the paperSupportsCheck (AI note)
Figs. S1–S2 (SI p.1)β turning into α at 230 °C; α-spherulite and α-needle side by side with the same Raman spectrum at 200 °C.·F2, F5
Figs. S3–S4 (SI p.2)γ turning into β at 190 °C; DSC at 100 °C/min showing β melting followed by recrystallization.·F5